Acylamino acid antihypertensives in the treatment of congestive heart failure

ABSTRACT

Mercapto-acylamino acids useful in the treatment of hypertension and combinations of mercapto-acylamino acids and atrial natriuretic factors or angiotensin converting enzyme inhibitors useful for treating hypertension are disclosed.

This application is a divisional of application Ser. No. 07/133,669,filed on Dec. 16, 1987, now U.S. Pat. No. 5,061,710, which in turn is acontinuation-in-part of Ser. No. 032,153, filed Mar. 27, 1987, now U.S.Pat. No. 4,801,609.

SUMMARY OF THE INVENTION

The present invention relates to mercapto-acylamino acids useful in thetreatment of hypertension and congestive heart failure.

The invention also relates to the treatment of hypertension andcongestive heart failure with a combination of a mercapto-acylamino acidand an atrial natriuretic factor (ANF) and with a combination of amercapto-acylamino acid and an angiotensin converting enzyme (ACE)inhibitor.

Other aspects of the invention relate to pharmaceutical compositionscomprising a mercapto-acylamino acid of this invention, alone or incombination with an ANF or an ACE inhibitor, and to methods of treatmentof hypertension and congestive heart failure comprising administering amercapto-acylamino acid of this invention, alone or in combination withan ANF or an ACE inhibitor to a mammal in need of such treatment.

BACKGROUND OF THE INVENTION

Human hypertension represents a disease of multiple etiologies. Includedamong these is a sodium and volume dependent low renin form ofhypertension. Drugs that act to control one aspect of hypertension willnot necessarily be effective in controlling another.

A variety of mercapto-acylamino acids are known as enkephalinaseinhibitors useful as analgesics and in the treatment of hypertension.

U.S. Pat. No. 4,513,009 to Roques et al discloses, inter alia, compoundsof the formula. ##STR1## wherein n is 0 to 1; R¹ includes hydrogen,optionally substituted alkyl, optionally substituted phenyl, cyclohexyland thienyl; and R² includes hydrogen optionally substituted alkyl,optionally substituted benzyl, phenyl, phenoxyalkyl and optionallysubstituted mercaptoalkyl. The compounds are disclosed as principallyhaving enkephalinase activity, but also are said to beantihypertensives.

U.S. Pat. No. 4,401,677 to Greenberg et al and EPA 38,046 to Wilkinsondisclose compounds of a scope similar to Roques et al, the formerdisclosing analgesic activity and the latter disclosing a greaterspecificity for enkephalinase than ACE. U.S. Pat. No. 4,053,651 toOndetti et al discloses the use of similar compounds in the treatment ofrenin-angiotensin related hypertension.

It has recently been discovered that the heart secretes a series ofpeptide hormones called atrial natriuretic factors (ANF) which help toregulate blood pressure, blood volume and the excretion of water, sodiumand potassium. ANF were found to produce a short-term reduction in bloodpressure and to be useful in the treatment of congestive heart failure.See P. Needleman et al, "Atriopeptin: A Cardiac Hormone IntimatelyInvolved in Fluid, Electrolyte and Blood-Pressure Homeostasis", N. Engl.J. Med., 314, 13 (1986) pp. 828-834, and M. Cantin et al in "The Heartas an Endocrine Gland", Scientific American, 254 (1986) pg. 76-81.

A class of drugs known to be effective in treating some types ofhypertension is ACE inhibitors, which compounds are useful in blockingthe rise in blood pressure caused by increases in vascular resistanceand fluid volume due to the formation of angiotensin II from Wyvratt andA. Patchett, "Recent Developments in the Design of AngiotensinConverting Enzyme Inhibitors" in Med. Res. Rev. Vol. 5, No. 4 (1985) pp.483-531.

DETAILED DESCRIPTION

Novel antihypertensive compounds of the present invention arerepresented by the following formulae: ##STR2## wherein R¹ is phenylsubstituted by one or more substituents independently selected fromalkyl, alkoxy, cycloalkyl, cyano and aminomethyl, Y--C₆ H₄ S--, Y--C₆ H₄O--, ##STR3## α-naphthyl, β-naphthyl, furyl, benzofuryl, benzothienyl,H₂ N(CH₂)m--, diphenylmethyl ##STR4## R² is alkyl, alkyl--S(O)₀₋₂(CH₂)_(q) --, R¹⁴ (CH₂)_(k) S(O)₀₋₂ (CH₂)_(q) --, alkyl-O(CH₂)_(q) --,R⁵ (CH₂)_(k) --O(CH₂)_(q) --, R⁵ (CH₂)_(q) --, H₂ N(CH₂)_(q) --,cycloalkyl(CH₂)_(k) --, R¹³ CONH(CH₂)_(q) --, R¹³ NHCO(CH₂)_(q) -- or R⁶OCO(CH₂)_(q) --; ##STR5## R³ is --OR⁷, R⁴ and R¹³ are independentlyhydrogen, alkyl or Y¹ --C₆ H₄ --;

R⁵ is Y² --C₆ H₄ --, Y² --C₆ H₄ S--, Y² --C₆ H₄ O--, α-naphthyl,β-naphthyl, furyl, thienyl, benzofuryl, benzothienyl, indolyl or##STR6## provided that when R⁵ is Y² --C₆ H₄ S-- or Y² --C₆ H₄ O--, k is2 or 3;

R¹⁴ is R⁵, mono-unsaturated lower alkyl, hydroxy, alkoxy or alkylthio,provided that when R¹⁴ is hydroxy or alkoxy, k is 2 or 3 and when R¹⁴ ismono-unsaturated alkykl or alkylthio, k is 1, 2 or 3;

R⁶, R⁷ and R⁸ are independently H, alkyl, hydroxyalkyl, dihydroxyalkyl,alkoxyalkyl, dialkoxyalkyl, alkoxyalkoxyalkyl, haloalkyl,(haloalkoxy)alkyl, aminoalkyl, alkylaminoalkyl, dialkylaminoalkyl,arylalkyl or alkyl substituted with a 5-6 membered saturated ringcomprising 1-2 oxygen atoms as ring members wherein the ring carbonatoms may be substituted with 0-2 alkyl substituents, or R⁷ and R⁸together with the nitrogen to which they are attached complete a 5-7membered ring, wherein one of the 4-6 ring members comprising R⁷ and R⁸may be a nitrogen atom, an alkyl-substituted nitrogen atom or an oxygenatom, and wherein the ring may be substituted on the ring carbon atomswith substituents chosen from alkyl and hydroxy groups;

R⁹ is hydrogen, alkyl, carboxyalkyl, mercaptoalkyl, alkylthioalkyl,aminoalkyl, hydroxyalkyl, phenylalkyl, hydroxyphenylalkyl,guanidinoalkyl, imidazolylalkyl, indolylalkyl or carbamoylalkyl;

n is 0-2;

m and k are independently 0-3;

q is 1-4;

X and X¹ are independently a bond, --O--, --S--, or --CH₂ --;

Q is hydrogen or R¹⁰ CO--;

R¹⁰ is alkyl, hydroxyalkyl, alkoxyalkyl, dialkylaminoalkyl, Y³ --C₆ H₄--alkyl, alkoxy, Y³ --C₆ H₄ --, naphthyl, furyl, thienyl or pyridyl;

Y, Y¹, Y² and Y³ independently represent one or more substituentsselected from H, alkyl, cycloalkyl, alkoxy, OH, F, Cl, Br, CN, --CH₂NH₂, --CO₂ H, --CO₂ alkyl, --CONH₂ and phenyl;

R^(1a) is Y--C₆ H₄ --, Y--C₆ H₄ S--, Y--C₆ H₄ O--, ##STR7## α-naphthyl,β-naphthyl, furyl, thienyl, benzofuryl, benzothienyl, H₂ N(CH₂)_(m) --,diphenylmethyl, ##STR8## R^(2a) is R^(14a) (CH₂)_(k) S(O)₀₋₂ (CH₂)_(q)--, R^(5a) (CH₂)_(k) --O (CH₂)_(q) --, R^(5a) (CH₂)_(q) --, orcycloalkyl-(CH₂)_(k), and when R³ is --NR⁷ R⁸, ##STR9## R^(2a) may alsobe indolyl--(CH₂)_(q) --, R¹³ CONH(CH₂)_(q) --, R¹³ NHCO(CH₂)_(q) -- orR⁶ OCO(CH₂)_(q) --; ##STR10## R^(3a) is R¹¹ is hydroxyalkyl,dihydroxyalkyl, alkoxyalkyl, dialkoxyalkyl, alkoxyalkoxyalkyl,haloalkyl, (haloalkoxy)alkyl, alkyl substituted with a 5-6 memberedsaturated ring comprising 1-2 oxygen atoms as ring members wherein thering carbon atoms may be substituted with 0-2 alkyl substituents, orsubstituted phenylalkyl wherein the phenyl group is substituted by oneor more groups selected from alkyl, alkoxy, cycloalkyl and cyano; R¹² isH or selected from the same group as R¹¹ ; or R¹¹ and R¹² together withthe nitrogen to which they are attached complete a 5-7 membered ringwherein one of the 4-6 ring members comprising R¹¹ and R¹² may be anitrogen atom, an alkyl-substituted nitrogen atom or an oxygen atom, andwherein the ring may be substituted on the ring carbon atoms withsubstituents chosen from alkyl and hydroxy groups;

R^(5a) is Y² --C₆ H₄ provided Y² is not H or OH, Y² --C⁶ H₄ S--, Y² --C₆H₄ O--, α-naphthyl, β-naphthyl, furyl, thienyl, benzofuryl, benzothienylor ##STR11## provided that when R^(5a) is Y² --C₆ H₄ --S-- or Y² --C₆ H₄O--, k is 2 or 3;

R^(14a) is R^(5a), mono-unsaturated alkyl, hydroxy, alkoxy or alkylthio,provided that when R^(14a) is hydroxy or alkoxy, k is 2 or 3 and whenR^(14a) is mono-unsaturated alkyl or alkylthio, k is 1, 2 or 3;

and the pharmaceutically acceptable addition salt thereof.

As used herein the term "alkyl" means straight or branched alkyl chainsof 1 to 6 carbon atoms, and "alkoxy" similarly refers to alkoxy groupshaving 1 to 6 carbon atoms. "Cycloalkyl" means cyclic alkyl groups of3-6 carbon atoms.

"Aryl" means mono-cyclic or fused ring bicyclic aromatic groups having 5to 10 ring members wherein 0-2 ring members may independently benitrogen, oxygen or sulfur and wherein the ring members may besubstituted by one to three substituents chosen from group Y definedabove. Examples of aryl groups are phenyl, α-naphthyl, β-naphthyl,furyl, thienyl, benzofuryl, benzothienyl, indolyl and pyridyl.

"Halo" refers to fluorine, chlorine, bromine or iodine radicals. Theterm "poly", when used to describe substitution in a phenyl, alkylphenylor alkoxyphenyl group, means 2 to 5 substituents.

Groups R³ and R^(3a) comprising the partial structure ##STR12## arederived from amino acids of formula ##STR13## Examples of such aminoacids are alanine, arginine, asparagine, aspartic acid, cysteine,glutamine, glutamic acid, glycine, histidine, isoleucine, leucine,lysine, methionine, phenylalanine, serine, threonine, tryptophan,tyrosine and valine.

Preferred embodiments of compounds of formula I are compounds wherein R²is alkyl, alkyl-S(O)₀₋₂ (CH₂)_(q) --, R¹⁴ (CH₂)_(k) S(O)₀₋₂ (CH₂)_(q) --or R⁵ (CH₂)_(q) --, wherein R¹⁴, R⁵, q and k are as defined above. Alsopreferred are compounds of formula I wherein R¹ is naphthyl, furyl,benzofuryl, benzothienyl, diphenylmethyl, aminoalkyl, Y--C₆ H₄ --X--C₆H₄ --, R⁴ CONH(CH₂)_(m) -- or R⁴ NHCO(CH₂)_(m) --, wherein Y, X, R⁴ andm are as defined above. A third group of preferred compounds in thatwherein R¹ is substituted phenyl. Still another group of preferredcompounds of formula I is that wherein R³ is --OR⁷ or --NR⁷ R⁸, whereinR⁷ and R⁸ are as defined above.

Preferred compounds of formula II are those wherein R^(2a) is R^(14a)(CH₂)_(k) S(O)₀₋₂ (CH₂)_(q) -- wherein R^(14a), q and k are as definedabove. Also preferred are compounds of formula II wherein R^(1a) isnaphthyl, furyl, thienyl, benzofuryl, benzothienyl, diphenylmethyl,aminoalkyl, Y-C₆ H₄ --X--C₆ H₄ --, R⁴ CONH(CH₂)_(m) -- or R⁴NHCO(CH₂)_(m) -- wherein Y, X, R⁴ and m are as defined above. A thirdgroup of preferred compounds is that wherein R^(1a) is Y--C₆ H₄. Stillanother group of preferred compounds of formula II is that wherein R³ is--OR⁷ or --NR⁷ R⁸, wherein R⁷ and R⁸ are as defined above.

Preferred compounds of formula III are those wherein R^(3a) is --OR¹¹,--NHCH₂ CONH₂, arylalkoxy or arylalkylamino. Also preferred arecompounds wherein R^(1a) is phenyl or substituted phenyl. Also preferredare compounds wherein R² is alkyl--S(O)₀₋₂ (CH₂)_(q) --, especially CH₃SCH₂ CH₂ -- or CH₃ SOCH₂ CH₂.

Preferred --OR⁷ and --OR¹¹ groups are those wherein the R⁷ and R¹¹groups are selected from (haloalkoxy)alkyl, alkoxyalkyl,alkoxyalkoxyalkyl, dihydroxyalkyl and2,2-dimethyl-1,3-dioxolan-4-yl-methyl.

Preferred compounds of formulae I-III are those wherein Q is hydrogen orR¹⁰ CO-- wherein R¹⁰ is alkyl, especially methyl, or phenyl.

Especially preferred are compounds of formula III wherein R^(1a) isoptionally substituted phenyl, n is 1, R² is CH₃ SCH₂ CH₂ -- or CH₃SOCH₂ CH₂ --, R^(3a) is --OR¹¹ wherein R¹¹ is alkoxyalkyl,dihydroxyalkyl, alkoxyalkoxyalkyl, (haloalkoxy)alkyl or2,2-dimethyl-1,3-dioxolan-4-yl-methyl, and Q is hydrogen, acetyl orbenzoyl.

Preferred compounds include the (R,S) and (S,S) isomers and thecorresponding free acids (i.e., R^(3a) is OH) of the following:

N-(2-benzoylthiomethyl-3-phenylpropionyl)-S-methionine2-(2-chloroethoxy)ethyl ester;

N-(2-benzoylthiomethyl-3-phenylpropionyl)-S-methionine 2-(ethoxy)ethylester;

N-(2-benzoylthiomethyl-3-phenylpropionyl)-S-methionine2,2-dimethyl-1,3-dioxolan-4-methyl ester;

N-(2-benzoylthiomethyl-3-phenylpropionyl)-S-methionine2-(2-methoxyethoxy)ethyl ester;

N-(2-acetylthiomethyl-3-phenylpropionyl)-S-methionine2-(2-chloroethoxy)ethyl ester;

N-(2-acetylthiomethyl-3-(4-methylphenyl)-propionyl)-S-methionine2-(2-chloroethoxy)ethyl ester;

N-(2-acetylthiomethyl-3-(2-methylphenyl)propionyl)-S-methionine2-(2chloroethoxy)ethyl ester;

N-(2-benzoylthiomethyl-3-phenylpropionyl)-S-methionine2,3-dihydroxypropyl ester; and

N-(2-benzoylthiomethyl-3-phenylpropionyl)-S-methionine sulfoxide2-(2-chloroethoxy)ethyl ester.

Compounds of this invention may, depending on the nature of functionalgroups, form addition salts with various inorganic and organic acids andbases. Such salts include salts prepared with organic and inorganicacids, e.g. HCl, HBr, H₂ SO₄, H₃ PO₄, methanesulfonic acid,toluenesulfonic acid, maleic acid, furmaric acid and camphorsulfonicacid. Salts prepared with bases include ammonium salts, alkali metalsalts, e.g. sodium and potassium salts, and alkaline earth salts, e.g.calcium and magnesium salts.

The salts may be formed by conventional means, as by reacting the freeacid or base forms of the product with one or more equivalents of theappropriate base or acid in a solvent or medium in which the salt isinsoluble, or in a solvent such as water which is then removed in vacuoor by freeze-drying or by exchanging the cations of an existing salt foranother cation on a suitable ion exchange resin.

Compounds of formulae I-III have two or more asymmetrical carbon atomsand therefore include various stereoisomers. All stereoisomers areincluded within the scope of the present invention.

Compounds of the present invention may be prepared by using couplingreactions well known in the peptide art to join a3-acetylthio-2-(substituted)propionic acid of formula 1 with an aminoacid ester of formula 2. The following reaction Scheme 1 is an example:##STR14##

In the above scheme, R^(p) =R¹ and R^(1a) ; R^(r) =R² and R^(2a) ; R^(t)is methyl, ethyl, t-butyl or aralkyl (e.g. benzyl); Ac is acetyl; n is0-2; DEC is 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride;HOBT is 1-hydroxybenzotriazole hydrate; NMM is N-methylmorpholine; andDMF is dimethylformamide.

As Scheme 1 shows, an amino acid ester of formula 2 and a 3-acetylthiopropionic acid of formula 1 are reacted at room temperature in an inertsolvent such as DMF in the presence of coupling agents such as DEC andHOBT in the presence of a base such as NMM. The resultant isomers areseparated by chromatography and the isomers are deprotected at the acidand mercapto positions.

Alternatively, a propionic acid of formula 1 may be reacted with thionylchloride to prepare the corresponding propionyl chloride, which may thenbe reacted with an amino acid ester of formula 2 or with thecorresponding free acid 2a in an inert solvent such as acetonitrile inthe presence of a base such as triethylamine to give isomers of formula3, which may be separated as in Scheme 1. The following Scheme 2 is anexample: ##STR15## wherein n, Ac, R^(p), R^(r) and R^(t) are as definedabove, and wherein R^(t) may also be hydrogen.

Other R³ and R^(3a) esters of compounds of formulae I-III are preparedby standard esterification techniques, for exampleN-(t-butoxycarbonyl)-S-methionine is reacted with2-(2-chloroethoxy)ethanol in the presence of a base such as4-dimethylaminopyridine, the amino function is deprotected and theresultant amino acid ester is reacted with a compound of formula 1 in amanner similar to that described in Scheme 2. In another example,N-(t-butoxycarbonyl)-S-methionine is reacted withN,N-diethylbromoacetamide and a reagent such as cesium carbonate, theresultant ester is deprotected at the amino function and a reactionsimilar to that described in Scheme 2 is carried out.

Compounds of formulae I-III wherein R³ or R^(3a) is --NR⁷ R⁸ areprepared by coupling reactions as described above in Schemes 1 and 2 byreplacing the amino acid ester 2 with an amide or substituted amide asshown in Scheme 3: ##STR16##

Alternatively, compounds of formulae I-III wherein R³ or R^(3a) is --NR⁷R⁸ may be prepared by coupling a propionyl chloride of formula 8 with anamino acid of formula 2a in the presence of a base and then coupling thedesired --NR⁷ R⁸ group to the carboxylic group using a typicalpeptide-coupling reaction. Scheme 4 shows an example of such aprocedure: ##STR17##

A third method for preparing compounds of formulae I-III wherein R³ orR^(3a) is --NR⁷ R⁸ comprises reacting a propionic acid of formula 1 withan amino acid t-butyl ester of formula 2, removing the t-butyl ester andcoupling the --NR⁷ R⁸ group to the carboxylic acid group as above.

Compounds wherein R³ and R^(3a) are ##STR18## are prepared analogouslyto those wherein R³ and R^(3a) are --NR⁷ R⁸.

Compounds wherein Q is R¹⁰ CO-- may be prepared by known methods, forexample by adding a mercaptoacid of formula R¹⁰ COSH to an acrylic acidto obtain a thio-substituted propionic acid analogous to compounds offormula 1. Alternatively, an amide of formulae I-III wherein Q is --SHmay be reacted with a compound of formulae R¹⁰ COCl is the presence of abase to obtain the desired sulfur substituted derivative.

Compounds wherein R² or R^(2a) is, e.g. alkyl-S(O)₁₋₂ (CH₂)_(q) -- orR¹⁴ (CH₂)_(k) --S(O)₁₋₂ (CH₂)_(q) --, are prepared by oxidizing withhydrogen peroxide the corresponding alkylthioalkyl compounds of formulaeI-III (e.g. those wherein R² or R^(2a) is, e.g. alkyl--S--(CH₂)_(q) --or R¹⁴ (CH₂)_(k) --S--(CH₂)_(q) --).

Starting materials of formulae 1 and 2 are known in the art or may beprepared by methods well known to those skilled in the art. Examples oftypical preparations of starting materials and specific examples ofcompounds of formulae I-III are provided at the end of thespecification.

A second aspect of the invention is the administration of a combinationof an ANF and a compound of the following formula IV ##STR19## whereinn, R^(1a), R², R³ and Q are as defined above, for the treatment ofhypertension. Compounds of formula IV have chiral centers and formaddition salts as described above for compounds of formulae I-III, andmay be prepared by similar methods.

As indicated by Needleman et al., a number of ANF have been isolated sofar, all having the same core sequence of 17 amino acids within acysteine disulfide bridge, but having different N-termini lengths. Thesepeptides represent N-terminal truncated fragments (21-48 amino acids) ofa common preprohormone (151 and 152 amino acids for man and rats,respectively). Human, porcine and bovine carboxy-terminal 28-amino acidpeptides are identical and differ from similar peptides in rats and micein that the former contain a methionine group at position 12 while thelatter contain isoleucine. Various synthetic analogs of naturallyoccurring ANFs also have been found to have comparable biologicalactivity. Examples of ANFs contemplated for use in this invention are αhuman AP 21 (atriopeptin I), α human AP 28, α human AP 23 (atriopeptinII or APII), α human AP 24, α human AP 25, α human AP 26, α human AP 33,and the corresponding rat sequence of each of the above wherein Met 12is Ile. See Table 1 for a comparison of the peptides.

    TABLE 1      ##STR20##      ##STR21##      ##STR22##      ##STR23##      ##STR24##      ##STR25##      ##STR26##     *Ile in the rat peptide

A third aspect of the invention in the administration of a combinationof an ACE inhibitor and a compound of formula IV.

Examples of ACE inhibitors are those disclosed in the article by Wyvrattet al., cited above, and in the following U.S. Pat. Nos. 4,105,776,4,468,519, 4,555,506, 4,374,829, 4,462,943, 4,470,973, 4,470,972,4,350,704, 4,256,761, 4,344,949, 4,508,729, 4,512,924, 4,410,520 and4,374,847, all incorporated herein by reference; and the followingforeign patents or published patent applications:

British Specification No. 2095682 published Oct. 6, 1982 disclosesN-substituted-N-carboxyalkyl aminocarbonyl alkyl glycine derivativeswhich are said to be angiotensin converting enzyme inhibitors and havethe formula ##STR27## either (A)

R^(b) and R₉ ^(b) are OH, 1-6C alkoxy, 2-6C alkenyloxy, di-(1-6Calkyl)amino-(1-6C) alkoxy, 1-6C hydroxyalkoxy, acylamino-(1-6C)alkoxy,acyloxy-(1-6C)alkoxy, aryloxy, aryloxy-(1-6C)alkoxy, NH₂, mono- ordi-(1-6C alkyl)amino, hydroxyamino or aryl-(1-6C)alkylamino; R₁ ^(b) -R₅^(b), R₇ ^(b) and R₈ ^(b) are 1-20C alkyl, 2-20C alkenyl, 2-20C alkynyl,aryl, aryl-(1-6C) alkyl having 7-12C or heterocyclyl-(1-6C)alkyl having7-12C;

R₆ ^(b) is cycloalkyl, polycycloalkyl, partly saturated cycloalkyl orpolycycloalkyl, cycloalkyl-(1-6C)alkyl having 3-20C, 6-10C aryl,aryl-(1-6C)alkyl, aryl-(2-6C)alkenyl or aryl-(2-6C) alkynyl; or

R₂ ^(b) and R₃ ^(b) together with the C and N atoms to which they areattached or R₃ ^(b) and R₅ ^(b) together with the N and C atoms to whichthey are attached form an N-heterocycle containing 3-5C or 2-4C and a Satom;

all alkyl, alkenyl and alkynyl are optionally substituted by OH, 1-6Calkoxy, thio(sic), 1-6C alkylthio, NH₂, mono- or di(1-6C alkyl)amino,halogen or NO₂ ;

all `cycloalkyl` groups (including poly and partially unsaturated) areoptionally substituted by halogen, 1-6C hydroxyalkyl, 1-6C alkoxy,amino-(1-6C alkyl)amino, di-(1-6C alkyl)amino, SH, 1-6C alkylthio, NO₂or CF₃ ; and

aryl groups are optionally substituted by OH, 1-6C alkoxy, NH₂, mono- ordi-(1-6C alkyl) amino, SH, 1-6C alkylthio, 1-6C hydroxyalkyl, 1-6Caminoalkyl, 1-6C thioalkyl, NO₂, halogen, CF₃, OCH₂ O, ureido orguanidino; or (B)

R^(b) and R₉ ^(b) are H or 1-6C alkoxy; R₁ ^(b) and R₂ ^(b) are H, 1-6Calkyl, aryl-(1-6C) alkyl having 7-12C or heterocyclyl-(1-6C) alkylhaving 6-12C;

R₃ ^(b) -R₅ ^(b), R₇ ^(b) and R₈ ^(b) are H or 1-6C alkyl; R₆ ^(b) iscycloalkyl, polycycloalkyl, partly saturated cycloalkyl orpolycycloalkyl, cycloalkyl-(1-6C) alkyl having 3-20C, aryl oraryl-(1-6C) alkyl; and

aryl has 6-10C and is optionally substituted by 1-6C alkyl, 2-6Calkenyl, 2-6C alkynyl, OH, 1-6C alkoxy, NH₂, mono- or di-(1-6C alkyl)amino, SH, 1-6C alkylthio, 1-6C hydroxyalkyl, 1-6C aminoalkyl, 1-6Cthioalkyl, NO₂, halogen, CF₃, OCH₂ O, ureido or guanidino;

European Patent Application 0 050 800 published May 5, 1982 disclosescarboxyalkyl dipeptides derivatives which are said to be angiotensinconverting enzyme inhibitors and have the formula ##STR28## or apharmaceutically acceptable salt thereof, wherein R^(c) and R^(6c) arethe same or different and are hydroxy, lower alkoxy, lower alkenyloxy,dilower alkylamino lower alkoxy, acylamino lower alkoxy, acyloxy loweralkoxy, aryloxy, aryllower alkoxy, amino, lower alkylamino, diloweralkylamino, hydroxyamino, aryllower alkylamino, or substituted aryloxyor substituted aryllower alkoxy wherein the substitutent is methyl, haloor methoxy; R^(1c) is hydrogen, alkyl of from 1 to 10 carbon atoms,substituted lower alkyl wherein the substitutent is hydroxy, loweralkoxy, aryloxy, substituted aryloxy, heteroaryloxy, substitutedheteroaryloxy, amino, lower alkylamino, diloweralkylamino, acylamino,arylamino, substituted arylamino, guanidino, imidazolyl, indolyl, loweralkylthio, arylthio, substituted arylthio, carboxy, carbamoyl, loweralkoxy carbonyl, aryl, substituted aryl, aralkyloxy, substitutedaralkyloxy, aralkylthio or substituted aralkylthio, wherein the aryl orheteroaryl portion of said substituted aryloxy, heteroaryloxy,arylamino, arylthio, aryl, aralkyloxy, aralkylthio group is substitutedwith a group selected from halo, lower alkyl, hydroxy, lower alkoxy,amino, aminomethyl, carboxyl, cyano, or sulfamoyl; R^(2c) and R^(7c) arethe same or different and are hydrogen or lower alkyl; R^(3c) ishydrogen, lower alkyl, phenyl lower alkyl, aminoethylphenyl lower alkyl,hydroxyphenyl lower alkyl, hydroxy lower alkyl, acylamino lower alkyl,amino lower alkyl, dimethylamino lower alkyl, guanidino lower alkyl,imidazolyl lower alkyl, indolyl lower alkyl, or lower alkyl thio loweralkyl; R^(4c) and R^(5c) are the same or different and are hydrogen,lower alkyl or Z^(c), or R^(4c) and R^(5c) taken together form a grouprepresented by Q^(c), U^(c), V^(c), Y^(c), D^(c) or E^(c), wherein;

Z^(c) is ##STR29## wherein X^(1c) and X^(2c) independent of each otherare O, S or CH₂, R^(8c) and R^(9c) independent of each other are loweralkyl, lower alkenyl, lower alkynyl, cycloalkyl having 3 to 8 carbonatoms, hydroxy lower alkyl, or --(CH₂)_(n) ^(c) Ar^(c), wherein n^(c) is0, 1, 2 or 3 and Ar^(c) is unsubstituted or substituted phenyl, furyl,thienyl or pyridyl, wherein said substituted phenyl, furyl, thienyl orpyridyl groups are substituted with at least one group that isindependently selected from C₁ to C₄ alkyl, lower alkoxy, loweralkylthio, halo, CF₃ and hydroxy, or R^(8c) and R^(9c) taken togetherform a bridge W^(c), wherein W^(c) is a single bond or a methylenebridge or a substituted methylene bridge when at least one of X^(1c) andX^(2c) is methylene, or W^(c) is an alkylene or substituted alkylenebridge having 2 or 3 carbon atoms, said substituted methylene bridge orsaid substituted alkylene bridge having one or two substituents selectedfrom lower alkyl, aryl and aryl lower alkyl groups, and p^(c) is 0, 1 or2; with the proviso that at least one of R^(4c) and R^(5c) is Z^(c),with the proviso that if R^(4c) is Z^(c) and p^(c) is 0 then X^(1c) andX^(2c) must both be methylene, and with the proviso that if X^(1c) andX^(2c) are both methylene then R^(8c) and R^(9c) must form an alkylenebridge W^(c) ;

Q^(c) is ##STR30## wherein R^(8c), R^(9c), X^(1c) and X^(2c) are asdefined above, p^(c) is 0, 1 or 2, q^(c) is 0, 1 or 2, with the provisothat the sum of p^(c) and q^(c) must be 1, 2 or 3, with the proviso thatif p^(c) is 0 then X^(1c) and X^(2c) must be methylene, and with theproviso that if X^(1c) and X^(2c) are methylene then R^(8c) and R^(9c)taken together form a bridge W^(c), wherein W^(c) is as defined above;

V^(c) is ##STR31## wherein R^(8c), R^(9c), X^(1c) and X^(2c) are asdefined above, p^(c) is 0, 1 or 2 and q^(c) is 0, 1 or 2, with theproviso that the sum of p^(c) and q^(c) is 1, 2 or 3, with the provisothat if X^(1c) and X^(2c) are CH₂ then R^(8c) and R^(9c) taken togetherform a bridge W^(c), wherein W^(c) is as defined above;

U^(c) is ##STR32## wherein W^(c) is as defined above (except that W^(c)may also be a methylene bridge when X^(1c) and X^(2c) are oxygen orsulfur), X^(1c) and X^(2c) are as defined above, p^(c) is 0, 1 or 2,q^(c) is 0, 1 or 2, with the proviso that the sum of p^(c) and q^(c) is1 or 2, and with the proviso that if p^(c) is 0, X^(1c) must be CH₂ ;Y^(c) is ##STR33## wherein G^(c) is oxygen, sulfur or CH₂, a^(c) is 2, 3or 4 and b^(c) is 1, 2, 3, 4 or 5, with the proviso that the sum ofa^(c) and b^(c) is 5, 6 or 7 or G^(c) is CH₂, a^(c) is 0, 1, 2 or 3,b^(c) is 0, 1, 2 or 3 with the proviso that the sum of ^(c) and b^(c) is1, 2 or 3, with the proviso that the sum of a^(c) and b^(c) may be 1, 2or 3 only if R^(1c) is lower alkyl substituted with aralkylthio oraralkyloxy; D^(c) is ##STR34## wherein F^(c) is O or S, j^(c) is 0, 1 or2 and k^(c) is 0, 1 or 2, with the proviso that the sum of j^(c) andk^(c) must be 1, 2 or 3, and m^(c) is 1, 2 or 3 and t^(c) is 1, 2 or 3,with the proviso that the sum of m^(c) and t^(c) must be 2, 3 or 4;

E^(c) is ##STR35## wherein L^(c) is O or S, u^(c) is 0, 1 or 2 and v^(c)is 0, 1 or 2, with the proviso that the sum of u^(c) and v^(c) must be 1or 2, and h^(c) is 1 or 2 and s^(c) is 1 or 2, with the proviso that thesum of h^(c) and s^(c) must be 2 or 3;

European Patent Application 0 079 522 published May 25, 1983 disclosesN-carboxymethyl(amidino) lysylproline compounds which are said to beangiotensin converting enzyme inhibitors and have the formula where##STR36## wherein: R^(d) and R^(2d) are independently hydrogen;loweralkyl; aralkyl; or aryl;

R^(1d) is hydrogen; branched or straight chain C₁₋₁₂ alkyl and alkenyl;C₃ -C₉ cycloalkyl and benzofused alkyl; substituted loweralkyl where thesubstituents are halo, hydroxy loweralkoxy, aryloxy, amino, mono- ordiloweralkylamino, acylamino, arylamino, guanidino, mercapto,loweralkylthio, arylthio, carboxy, carboxamido, or loweralkoxycarbonyl;aryl; substituted aryl where the substituents are loweralkyl,loweralkoxy, or halo; arloweralkyl; arloweralkenyl; heteroarloweralkyl;heteroarloweralkenyl; substituted arloweralkyl, substitutedarloweralkenyl, substituted heteroarloweralkyl, or substitutedheteroarloweralkenyl where the aryl and heteroaryl substituents arehalo, dihalo, loweralkyl, hydroxy, loweralkoxy, amino, aminoloweralkyl,acylamino, mono- or diloweralkylamino, carboxyl, haloloweralkyl, nitro,cyano, or sulfonamido, and where the loweralkyl portion of arloweralkylmay be substituted by amino, acylamino, or hydroxyl; ##STR37## where:

X^(d) and Y^(d) taken together are --CH₂ --CH₂ --; ##STR38## R^(4d) ishydrogen; loweralkyl; aryl or substituted aryl; R^(5d) is hydrogen;loweralkyl; aryl or substituted aryl;

n^(d) is 1 to 3;

W^(d) is absent; ##STR39## Z^(d) is --(CH₂)_(m) ^(d), where m^(d) is 0to 2, provided that m^(d) may not be 0 and W^(d) may not be absent atthe same time; and

R^(6d) is hydrogen; loweralkyl; halo; or OR^(4d) ;

R^(2d) is --(CH₂)_(r) ^(d) --B^(d) --(CH₂)_(s) ^(d) --NR^(7d) R^(15d)

where

r^(d) and s^(d) are independently 0 to 3;

B^(d) is absent; --O--; --S--; or --NR^(8d) ;

where R^(8d) is hydrogen; loweralkyl; alkanoyl; or aroyl; and

R^(7d) is, ##STR40## where R^(9d) is loweralkyl; aralkyl; aryl;heteroaryl; or heteroarloweralkyl and these groups substituted byhydroxy, lower alkoxy or halo; carboxyl; carboxamido; nitromethenyl.

R^(10d) is hydrogen; loweralkyl; aryl; or amidino;

R^(11d) hydrogen; loweralkyl; cyano; amidino; aryl; aroyl;loweralkanoyl; ##STR41## --NO₂ ; --SO₂ NH₂ ; or SO₂ R^(13d) ; R^(12d) ishydrogen; loweralkyl; halo; aralkyl; amino; cyano; mono- ordiloweralkylamino; or OR^(4d) ;

R^(13d) is hydrogen; loweralkyl; or aryl;

R^(15d) is hydrogen; lower alkyl; aralkyl; or aryl; ##STR42## constitutea basic heterocycle of 5 or 6 atoms or benzofused analogs thereof andoptionally containing 1-3N atoms, an oxygen, a sulfur, an S═O, or an SO₂group optionally substituted by amino, lower alkyl amino, diloweralkylamino, lower alkoxy, or aralkyl groups;

R^(3d) is C₃₋₈ cycloalkyl and benzofused C3-8 cycloalkyl;perhydrobenzofused C3-8 cycloalkyl; aryl; substituted aryl; heteraryl;substituted heteroaryl;

R^(14d) is hydrogen or loweralkyl; and, a pharmaceutically acceptablesalt thereof;

European Patent 79022 published May 18, 1983 discloses N-aminoacyl-azabicyclooctane carboxylic acid derivatives which have the formula##STR43## wherein hydrogen atoms at ring positions 1 and 5 are cis toeach other and the 3-carboxy group has the endo orientation;

R₁ ^(e) is H, allyl, vinyl or the side chain of an optionally protectednaturally occurring α-amino acid;

R₂ ^(e) is H, 1-6C alkyl, 2-6C alkenyl or aryl(1-4C alkyl);

Y^(e) is H or OH and Z^(e) is H, or Y^(e) and Z^(e) together oxygen;

X^(e) is 1-6C alkyl, 2-6C alkenyl, 5-9C cycloalkyl, 6-12C aryl(optionally substituted by one to three 1-4C alkyl or alkoxy, OH, halo,nitro, amino (optionally substituted by one or two 1-4C alkyl), ormethylenedioxy) or indol-3-yl);

European Patent 46953 published Mar. 10, 1982 discloses N-aminoacyl-indoline and tetrahydro isoquinoline carboxylic acids which areangiotensin coverting enzyme inhibitors and have the formula ##STR44##n^(f) is 0 or 1; ##STR45##

is a benzene or cyclohexane ring: R₁ ^(f) and R₂ ^(f) are each 1-6Calkyl, 2-6C alkenyl, 5-7C cycloalkyl, 5-7C cycloalkenyl, 7-12Ccycloalkylalkyl, optionally partially hydrogenated 6-10C aryl, 7-14Caralkyl or 5-7 membered monocyclic or 8-10 membered bicyclicheterocyclyl containing 1 or 2 S or O and/or 1-4N atoms; all R₁ ^(f) andR₂ ^(f) groups are optionally substituted, R₃ ^(f) is H, 1-6C alkyl,2-6C alkenyl or 7-14C aralkyl.

The following Table I lists ACE inhibitors preferred for use in thecombination of this invention.

                                      TABLE I                                     __________________________________________________________________________    PREFERRED ACE INHIBITORS                                                      __________________________________________________________________________     ##STR46##                                                                           R         R.sub.1                                                                          R.sub.2 R.sub.3                                           __________________________________________________________________________    spirapril                                                                            C.sub.6 H.sub.5 CH.sub.2 CH.sub.2                                                       Et CH.sub.3                                                                               ##STR47##                                        enalapril                                                                            C.sub.6 H.sub.5 CH.sub.2 CH.sub.2                                                       Et CH.sub.3                                                                              prolyl                                            ramipril                                                                             C.sub.6 H.sub.5 CH.sub.2 CH.sub.2                                                       Et CH.sub.3                                                                               ##STR48##                                        perindopril                                                                          CH.sub.3 CH.sub.2 CH.sub.2                                                              Et CH.sub.3                                                                               ##STR49##                                        indolapril                                                                           C.sub.6 H.sub.5 CH.sub.2 CH.sub.2                                                       Et CH.sub.3                                                                               ##STR50##                                        lysinopril                                                                           C.sub.6 H.sub.5 CH.sub.2 CH.sub.2                                                       H  NH.sub.2 (CH.sub.2).sub.4                                                             prolyl                                            CI-925 C.sub.6 H.sub.5 CH.sub.2 CH.sub.2                                                       Et CH.sub.3                                                                               ##STR51##                                        pentopril (NH = CH.sub.2)                                                            CH.sub.3  Et CH.sub.3                                                                               ##STR52##                                        cilazapril                                                                           C.sub.6 H.sub.5 CH.sub.2 CH.sub.2                                                       H                                                                                 ##STR53##                                                                             ##STR54##                                        __________________________________________________________________________     ##STR55##                                                                                 R             R.sub.2                                            __________________________________________________________________________    captopril    H             prolyl                                             zofenopril   C.sub.6 H.sub.5 CO                                                                           ##STR56##                                         pivalopril                                                                                  ##STR57##                                                                                   ##STR58##                                         __________________________________________________________________________     ##STR59##                                                                           R          R.sup.1       R.sup.2                                       __________________________________________________________________________    fosinopril                                                                           C.sub.6 H.sub.5 (CH.sub.2).sub.4                                                          ##STR60##    C.sub.6 H.sub.5                               __________________________________________________________________________

We have found that the novel compounds of the present invention areeffective in treating congestive heart failure and various types ofhypertension, particularly volume expanded hypertension. These novelcompounds as well as other mercapto-acylamino acids known in the arthave been found to enhance both the magnitude and duration of theantihypertensive and natriuretic effects of endogenous ANF.Administration of a combination of a mercapto-acylamino acid and an ACEinhibitor provides an antihypertensive effect greater than either themercapto-acylamino acid or ACE inhibitor alone. Administration of acombination of a mercapto-acylamino acid of formula IV and an exogenousANF or ACE inhibitor is therefore particularly useful in treatinghypertension.

In addition to the compound aspect, the present invention therefore alsorelates to treating hypertension with a mercapto-acylamino acid or witha mercapto-acylamino acid in combination with an ANF or an ACEinhibitor, which methods comprise administering to a mammal in need ofsuch treatment an antihypertensive effective amount of themercapto-acylamino acid or an antihypertensive effective amount of acombination of a mercapto-acylamino acid and ANF or ACE inhibitor. Thedrug or combination of drugs is preferably administered in apharmaceutically acceptable carrier, e.g. for oral or parenteraladministration. The combinations of drugs may be co-administered in asingle composition, or components of the combination therapy may beadministered separately. Where the components are administeredseparately, any convenient combination of dosage forms may be used, e.g.oral mercapto-acylamino acid/oral ANF, oral mercapto-acylaminoacid/parenteral ACE inhibitor, parenteral mercapto-acylamino acid/oralANF, parenteral mercapto-acylamino acid/parenteral ACE inhibitor.

When the components of a combination of a mercapto-acylamino acid and anANF are administered separately, it is preferred that themercapto-acylamino acid be administered first.

The present invention also relates to a pharmaceutical compositioncomprising a mercapto-acylamino acid for use in treating hypertension,to a pharmaceutical composition comprising both a mercapto-acylaminoacid and an ANF and to a pharmaceutical composition comprising both amercapto-acylamino acid and an ACE inhibitor.

The antihypertensive effect of mercapto-acylamino acids alone and incombination with ACE inhibitors was determined according to thefollowing procedure:

Male Sprague Dawley rats weighing 100-150 g were anesthetized with etherand the right kidney was removed. Three pellets containing Doc acetate(desoxycorticosterone acetate, DOCA, 25 mg/pellet) were implantedsubcutaneously. Animals recovered from surgery, were maintained onnormal rat chow and were allowed free access to a fluid of 1% NaCl and0.2% KCl instead of tap water for a period of 17-30 days. This procedureresults in a sustained elevation in blood pressure and is a slightmodification of published procedures (e.g. Brock et al., 1982) that havebeen used to produce DOCA salt hypertension in the rat.

On the day of study, animals were again anesthetized with ether and thecaudal artery was cannulated for blood pressure measurement. Patency ofthe caudal artery cannula was maintained with a continuous infusion ofdextrose in water at a rate of 0.2 ml/hr. Animals were placed intorestraining cages where they recovered consciousness. Blood pressure wasmeasured from caudal artery catheter using a Statham pressure transducerattached to a Beckman oscillographic recorder. In addition, acardiovascular monitoring device (Buxco Electronics, Inc.) and a digitalcomputer were used to calculate average blood pressures.

After an equilibration period of at least 1.5 hr., animals were dosedsubcutaneously (1 ml/kg) with vehicle (methylcellulose, hereinafter MC)or mercapto-acylamino acid and blood pressure was monitored for the next4 hours.

The antihypertensive effect of mercapto-acylamino acids in combinationwith ANF was determined according to the following procedures:

Male spontaneously hypertensive rats (SHR), 16-18 weeks old, 270-350 g,were anesthetized with ether and the abdominal aorta was cannulatedthrough the tail artery. The animals were then placed into restrainersto recover from anesthesia (in less than 10 min.) and remained insidethroughout the experiments. Through a pressure transducer (Gould P23series) analog blood pressure signals were registered on a Beckman 612recorder. A Buxco digital computer was used to obtain mean arterialpressures. Patency of the arterial cannula was maintained with acontinuous infusion of 5% dextrose at 0.2 ml/hr. Animals were allowed a90-min equilibration period. The animals first underwent a challengewith an ANF such as atriopeptin II (AP II) or AP28 30 μg/kg iv and atthe end of 60 min. were treated with drug vehicle or amercapto-acylamino acid subcutaneously. A second ANF challenge wasadministered 15 min. later and blood pressure was monitored for the next90 min.

The antihypertensive effect in SHR of mercapto-acylamino acids and ACEinhibitors, alone and in combination, was determined as follows:

Animals were prepared for blood pressure measurement as described above.After stabilization, animals were dosed subcutaneously or orally withtest drugs or placebo and blood pressure was monitored for the next 4hr.

The compositions of this invention comprise a mercapto-acylamino acid ora mercapto-acylamino acid and an ANF or a mercapto-acylamino acid and anACE inhibitor in combination with a pharmaceutically acceptable carrierfor administration to mammals. A variety of pharmaceutical forms issuitable, preferably for oral or parenteral administration, althoughmechanical delivery systems such as transdermal dosage forms are alsocontemplated.

The daily antihypertensive dose of the compound or combinations of thisinvention is as follows: for mercapto-acylamino acids alone the typicaldosage is 1 to 100 mg/kg of mammalian weight per day administered insingle or divided dosages; for the combination of mercapto-acylaminoacid and an ANF, the typical dosage is 1 to 100 mg of mercapto-acylaminoacid/kg mammalian weight per day in single or divided dosages plus 0.001to 0.1 mg ANF/kg of mammalian weight per day, in single or divideddosages, and for the combination of mercapto-acylamino acid and an ACEinhibitor, the typical dosage is 1 to 100 mg of mercapto-acylaminoacid/kg mammalian weight per day in single or divided dosages plus 0.1to 30 mg ACE inhibitor/kg of mammalian weight per day in single ordivided dosages. The exact dose of any component or combination to beadministered is determined by the attending clinician and is dependenton the potency of the compound administered, the age, weight, conditionand response of the patient.

Generally, in treating humans having hypertension, the compounds orcombinations of this invention may be administered to patients in adosage range as follows: for treatment with mercapto-acylamino acidsalone, about 10 to about 500 mg per dose given 1 to 4 times a day,giving a total daily dose of about 10 to 2000 mg per day; for thecombination of mercapto-acylamino acid and ANF, about 10 to about 500 mgmercapto-acylamino acid per dose given 1 to 4 times a day and about0.001 to about 1 mg ANF given 1 to 6 times a day (total daily dosagerange of 10 to 2000 mg day and 0.001 to 6 mg/day, respectively); and forthe combination of a mercapto-acylamino acid and an ACE inhibitor, about10 to about 500 mg mercapto-acylamino acid per dose given 1 to 4 times aday and about 5 to about 50 mg ACE inhibitor given 1 to 3 times a day(total daily dosage range of 10 to 2000 mg/day and 5 to 150 mg/day,respectively). Where the components of a combination are administeredseparately, the number of doses of each component given per day may notnecessarily be the same, e.g. where one component may have a greaterduration of activity, and will therefore need to be administered lessfrequently.

Typical oral formulations include tablets, capsules, syrups, elixirs andsuspensions. Typical injectable formulations include solutions andsuspensions.

The typical acceptable pharmaceutical carriers for use in theformulations described above are exemplified by: sugars such as lactose,sucrose, mannitol and sorbitol, starches such as cornstarch, tapiocastarch and potato starch; ceullulose and derivatives such as sodiumcarboxymethyl cellulose, ethyl cellulose and methyl cellulose; calciumphosphates such as dicalcium phosphate and tricalcium phosphate; sodiumsulfate; calcium sufate; polyvinylpyrrolidone, polyvinyl alcohol;stearic acid; alkaline earth metal stearates such as magnesium stearateand calcium stearate, stearic acid, vegetable oils such as peanut oil,cottonseed oil, sesame oil, olive oil and corn oil; non-ionic, cationicand anionic surfactants; ethylene gylcol polymers; betacyclodextrin;fatty alcohols and hydrolyzed cereal solids; as well as other nontoxiccompatible fillers, binders, disintegrants, buffers, preservatives,antioxidants, lubricants, flavoring agents, and the like commonly usedin pharmaceutical formulations.

Since the present invention relates to treatment of hypertension with acombination of active ingredients wherein said active ingredients may beadministered separately, the invention also relates to combiningseparate pharmaceutical compositions in kit form. That is, two kits arecontemplated, each combining two separate units: a mercapto-acylaminoacid pharmaceutical composition and an ANF pharmaceutical composition inone kit and a mercapto-acylamino acid pharmaceutical composition and anACE inhibitor pharmaceutical composition in a second kit. The kit formis particularly advantageous when the separate components must beadministered in different dosage forms (e.g. oral and parenteral) or areadministered at different dosage intervals.

Following are descriptions of preparations of typical starting materialsand examples of procedures for preparing compounds of formulae I-IV.Temperature designations, i.e. reaction conditions and melting points,are in ° C.

Preparation 1 L-Cysteine Esters Method 1 S-(4-Methylbenzyl)-L-Cysteine,Methyl Ester, Hydrochloride

At room temperature, add thionyl chloride (2.8 ml, 2.2 equiv.) dropwiseto N-t-butyloxycarbonyl-S-(4-methylbenzyl)-L-cysteine (5.0 g) inmethanol (500 ml) and heat the resulting mixture under reflux for 90minutes. Cool the reaction mixture to room temperature and concentratein vacuo to give the title compound, a white solid (4.31 g), m.p.158°-160°, [α]_(D) ²⁶ =-22.9° (MeOH).

By the same method, other amino acid esters are prepared:

S-Benzyl-D-cysteine ethyl ester hydrochloride, a while solid, m.p.149°-151°, [α]_(D) ²⁶ =+15.5° (H₂ O);

S-(4-Methoxybenzyl)-L-cysteine methyl ester hydrochloride, a whitesolid, m.p. 145°-6°, [α]_(D) ²⁶ =-23.2° (MeOH);

S-(3,4-Dimethylbenzyl)-L-cysteine ethyl ester hydrochloride, whitesolid, m.p. 161°-167°, [α]_(D) ²⁶ =-26.6° (MeOH); and

S-t-Butyl-L-cysteine methyl ester, an oil.

Method 2 S-Phenethyl-L-Cysteine Ethyl Ester Hydrochloride

Add thionyl chloride (2.0 ml) to absolute ethanol (25 ml) at 0°14 5°. Tothis solution add S-phenethyl-L-cysteine (2.5 g, 1.11 mmole). Warm theresulting mixture to room temperature, and then heat at 60° for 5 hours.Concentrate the reaction mixture in vacuo, dissolve the resultantresidue in dichloromethane (CH₂ Cl₂), and concentrate the solution invacuo. Dissolve the residue in absolute ethanol, treat with DARCO,filter and concentrate in vacuo to give the title compound, a whitesolid (2.90 g) m.p. 155°-156°, [α]_(D) ²⁶ =-1.5° (MeOH).

By the same method, other amino acid esters are prepared:

O-Benzyl-L-tyrosine methyl ester hydrochloride, a white solid, m.p.189-190, [α]_(D) ²⁶ =+6.9° (MeOH);

S-Methyl-L-cysteine ethyl ester, an oil, [α]_(D) ²⁶ =+25.1° (MeOH);

S-Ethyl-L-cysteine ethyl ester hydrochloride, a white solid, m.p.130°-3° [α]_(D) ²⁶ =-11.0° (MeOH); and

Ethionine ethyl ester, hydrochloride, clear oil, [α]_(D) ²⁶ =+15.5°(MeOH).

Preparation 2 3-Acetylthio-2-(Arylmethyl)Propionic Acids3-Acetylthio-2-(4-phenylbenzyl)propionic acid Step 1

4-Biphenylmethylenemalonic acid: Heat 4-biphenyl carboxaldehyde (18.0 g,9.89 mmole) and malonic acid (10.8 g, 10.4 mmole) in glacial acetic acid(6 ml) at approximately 100° for 2 hours. Cool the reaction mixture,dilute with dichloromethane and filter to give a white solid (8.76 g).Concentrate the filtrate, add malonic acid (1.5 g) and heat theresulting mixture at 100° for 2 hours. Cool the reaction mixture, dilutewith CH₂ Cl₂ and filter to give a white solid (3.44 g). Suspend thecombined solids (12.20 g) in water and filter to give the titlecompound, a pale yellow solid (9.56 g), m.p. 208°-9°↑.

By this method other aryl methylene malonic acids are prepared, forexample:

β-Naphthylmethylene malonic acid, a white solid, m.p. 204°-205°.

Step 2

(4-Phenylbenzyl)malonic acid: Hydrogenate the product of Step 1 (9.50 g,3.5 mmole) in ethyl acetate (200 ml) in the presence of 10%palladium-on-charcoal (0.80 g) at 50 psi for 3 hours. Filter andconcentrate the reaction mixture in vacuo to give the title compound, awhite solid (8.16 g) m.p. 180°-181°.

By this method other arylmethyl malonic acids are prepared, for example:

β-Naphthylmethyl malonic acid, a white solid, m.p. 150°-152°.

Step 3

2-(4-Phenylbenzyl)acrylic acid: Treat a portion of the product of Step 2(4.05 g, 1.50 mmole) in water (20 ml) at 0°-5° with 40% dimethylamine inwater to pH 7-8. Add the remaining product of Step 2 (4.05 g, 1.50mmole). After 15 minutes, add aqueous formaldehyde (10.0 ml, 38%).Slowly warm the resulting mixture to room temperature and stir for 18hours. Filter the reaction mixture, wash the white solid with water, andsuspend the solid in water (150 ml). Heat this suspension at 100° for 3hours until the solution is almost clear. Cool the solution and addconcentrated hydrochloric acid to pH 2 to give a white precipitate.Filter the mixture and dry the white solid. Dissolve this white solid inhot methanol, filter and concentrate in vacuo to give the titlecompound, a white solid (6.68 g) m.p. 168°-170°.

By this method other 2-(aryl)acrylic acids are prepared, for example:

2-(β-naphthylmethyl)acrylic acid, a white solid, m.p. 83°-84°.

Step 4

3-Acetylthio-2-(4-phenylbenzyl)propionic acid Add thioacetic acid (8.0ml) to the product of Step 3 (6.0 g, 2.77 mmole) in CH₂ Cl₂ (30 ml) andethyl acetate (100 ml) and stir the resulting mixture at roomtemperature for 72 hours. Concentrate the reaction mixture in vacuo.Dissolve the residue in toluene (100 ml), and concentrate in vacuo (3times) to give a yellow oil (6.0 g). Chromatograph the oil on a columnof silica gel (1.6L), eluting with CH₂ Cl₂ (4L) CH₂ Cl₂ :methanol 1000:1(3L), and 1000:5 (15L) to give the title compound, a white solid (3.44g), m.p. 101°-103°.

By this method, other 3-acetylthio-2-(arylmethyl)propionic acids areprepared:

3-Acetylthio-2-(4-chlorobenzyl)propionic acid, an oil;

3-Acetylthio-2-(α-naphthylmethyl)propionic acid, a white solid, m.p.94°-7°;

3-Acetylthio-2-(β-naphthylmethyl)propionic acid, a white solid, m.p.103°-106°;

3-Acetylthio-2-(4-methylbenzyl)propionic acid, m.p. 72°-75°; and

3-Acetylthio-2-(2-methylbenzyl)propionic acid, a waxy solid.

PREPARATION 3 1-(α-Naphthylmethyl)Acrylic Acid Step 1

Diethyl α-naphthylmethyl malonate: Add sodium metal (11.0 g, 0.478 mole)to absolute ethanol (650 ml) with cooling and stirring until the sodiumis dissolved. Add diethyl malonate (75.6 g, 0.473 mole) over 15 minutesat room temperature. After 30 min., add α-bromomethylnaphthalene (100 g,0.452 mole) in absolute ethanol (400 ml). Heat the resulting mixtureunder reflux for 5 hours. Keep at room temperature for 20 hours, andconcentrate in vacuo. Partition the residue between water (500 ml) anddiethyl ether (700 ml). Extract the diethyl ether solution with water(200 ml), and brine (200 ml), then dry the organic layer (MgSO₄) andconcentrate in vacuo to give the title compound, an oil (133.7 g).

Step 2

3-(1-Naphthyl)-2-ethoxycarbonylpropionic acid: To the product of Step 1(133.7 g, 0.446 mole) in absolute ethanol (400 ml) add a solution ofpotassium hydroxide (24.9 g, 0.445 mole) in absolute ethanol (400 ml)and stir the resulting mixture at room temperature for 20 hours.Concentrate the reaction mixture in vacuo and partition the residuebetween ice water (1L) and diethyl ether (500 ml). Cool the aqueoussolution to 0°-5° C. and acidify to approximately pH 2 with 2Nhydrochloric acid, keeping the temperature at 0°-5°. Extract the mixturewith diethyl ether, dry the organic layer (MgSO₄) and concentrate invacuo to give the title compound, an oil (100 g).

Step 3

Ethyl 1-(α-naphthylmethyl)acrylate: Add to the product of Step 2 (100 g,0.367 mole) and diethylamine (39 ml) a 37% aqueous solution offormaldehyde (38 ml) over 30 min. at 0°-5° C. with vigorous stirring.Stir the mixture at room temperature for 7 hours and then extract withdiethyl ether (3×500 ml). Extract the organic layer with 2N hydrochloricacid (2×500 ml), saturated aqueous sodium bicarbonate solution (500 ml)and brine (500 ml). Dry the organic layer (MgSO₄) and concentrate invacuo to give the title compound, an oil (58.6 g).

Step 4

1-(α-Naphthylmethyl)acrylic acid: Treat the product of Step 3 (12.0 g,50 mmole) in dioxane (50 ml) with 1N sodium hydroxide (60 ml) and stirthe resulting mixture at room temperature for 18 hours. Concentrate thereaction mixture under nitrogen, dilute with water and extract withethyl acetate. Cool the aqueous solution to 0°-5° and add concentratedhydrochloric acid slowly to pH 3-4 to give a white solid. Filter thereaction mixture, wash the resultant white solid with water and dry togive the title compound, a white solid (9.62 g), m.p. 115°-117°.

PREPARATION 4 L-Bishomophenylalanine t-Butyl[(2S)-Amino-5-Phenylpentanoic Acid, t-Butyl Ester] Step 1

4-Benzoyl-2(S)-trifluoroacetamidobutyric acid: Heat at reflux for 3hours a mixture of N-trifluoroacetyl-L-glutamic anhydride (18.0 g, 80mmol) and AlCl₃ (23.5 g, 177 mmol) in dry benzene (400 ml). Allow tocool, treat with ice (400 ml), conc. HCl (100 ml), and ethyl acetate(EtOAc) (400 ml). Dry the organic layer and concentrate to obtain thetitle compound as a brown crystalline solid (25 g).

Step 2

5-Phenyl-2(S)-trifluoroacetamidopentanoic acid: Reduce the product ofStep 1 (9.7 g) with Pearlman's catalyst (3.5 g) in EtOAc (75 ml) andethanol (25 ml) at 50 psi H₂ for 3 hours. Filter, concentrate and washthe resultant residue with 3:1 hexane:diethyl ether to give the titlecompound (7.8 g).

Step 3

N-Trifluoroacetyl-L-bishomophenylalanine, t-butyl ester: Treat theproduct of Step 2 (11.3 g) with isobutylene (25 ml) and conc. H₂ SO₄(1.0 ml) in CH₂ Cl₂ (100 ml) for 16 hours. Partition between diethylether and 1N NaHCO₃, dry, concentrate, and chromatograph the resultantresidue on silica gel, eluting with 2:1 hexane:diethyl ether to obtain11.3 g of the title compound as a colorless oil.

Step 4

L-Bishomophenylalanine, t-butyl ester, hydrochloride: To the product toStep 3 in EtOH (120 ml) add NaBH₄ (5.2 g) portionwise over 30 min. Stiranother 1.5 hours, concentrate, and partition between diethyl ether andH₂ O. Dry and concentrate to obtain an oil (8.1 g). Treat withHCl:diethyl ether to give the hydrochloride salt of the title compound,white crystals (4.0 g), m.p. 161°-2°, [α]_(D) ²⁶ =+15.6° (MeOH, c=0.5).

PREPARATION 5 S-(4-Methylbenzyl)-L-Cysteine Amide Step 1

N-t-Butyloxycarbonyl-S-(4-methylbenzyl)-L-cysteine amide: ReactN-t-butyloxycarbonyl-S-(4-methylbenzyl)-L-cysteine (6.50 g) withtriethylamine (4.44 g, 6.16 ml) in tetrahydrofuran (THF). Cool themixture to 0°-5°. Add ethyl chloroformate (4.77 g, 3.41 ml) in THF (5ml) dropwise over 5 min. and stir the reaction mixture for 15 min. Addammonium hydroxide (28%, 2.0 ml) in THF (5 ml) dropwise. Allow thereaction mixture to warm to room temperature and stir for 18 hr. Filterthe reaction mixture and concentrate the filtrate in vacuo to give apale yellow solid. Dissolve this solid in CH₂ Cl₂ and extract with H₂ O.Concentrate the dried (MgSO₄) CH₂ Cl₂ solution in vacuo to give a paleyellow solid (6.41 g). Recrystallize this solid from EtOAc to give thetitle compound, a white solid (2.82 g), m.p. 140°-141°, [α]_(D) ²⁶=-7.5° (MeOH).

Step 2

S-(4-Methylbenzyl)-L-cysteine amide: Treat the product of Step 1 (2.79g) in CH₂ Cl₂ (40 ml) with trifluoroacetic acid (10 ml) at roomtemperature for 18 hr. Concentrate the reaction mixture in vacuo.Dissolve the residue in CH₂ Cl₂ and concentrate in vacuo (twice).Dissolve the white solid in EtOAc and extract with 10% sodiumbicarbonate solution. Dry (MgSO₄) the EtOAc and concentrate in vacuo togive the title compound, a white solid (1.53 g) m.p. 94°-95°, [α]_(D) ²⁶=-1.3° (MeOH).

PREPARATION 6 S-(4-Methylbenzyl)-L-Cysteine t-Butyl Ester

To a cold solution of isobutylene (50 ml) in dioxane (80 ml), addS-(4-methylbenzyl)-L-cysteine (5.0 g) and concentrated H₂ SO₄ (10 ml).Seal the vessel, allow to warm to room temperature, and stir for 18 hr.Pour into 5% NaOH (500 ml), extract with Et₂ O (3×400 ml), dry (MgSO₄)and concentrate the Et₂ O in vacuo to give an oil. Treat the oil withHCl in Et₂ O to give the title compound, white needles (2.76 g) m.p.218° (dec).

EXAMPLE 1 N-(2-Benzyl-3-Mercaptopropionyl)-S-(4-Methylbenzyl)-L-Cysteine(Isomers A and B) Step 1:N-(3-Acetylthio-2-Benzylpropionyl)-S-(4-Methylbenzyl)-L-Cysteine MethylEster (Isomers A & B)

Add S-(4-methylbenzyl)-L-cysteine methyl ester, hydrochloride (1.85 g,0.77 mmole) to 3-acetylthio-2-benzyl propionic acid (1.93 g, 0.81mmole), DEC (1.46 g, 0.76 mmole), HOBT (1.18 g, 0.76 mmole) and NMM(2.30 g, 2.5 ml, 2.27 mmole) in DMF (20 ml) and stir the resultingmixture at room temperature for 20 hours. Concentrate the reactionmixture in vacuo and partition the residue between EtOAc and water.Concentrate the dried (MgSO₄) ethyl acetate solution in vacuo to give anamber oil (3.85 g). Chromatograph the oil on silica gel (Baker, 60-200mesh) (1.5L) using EtOAc:hexane 4:21 as eluent to give Isomer A, whitesolid (0.64 g), m.p. 101°-104°, [α]_(D) ²⁶ =-76.2° (MeOH); overlapIsomer A and Isomer B (0.40 g); and Isomer B, white solid (0.67 g), m.p.52°-55° [α]_(D) ²⁶ =- 4.9° (MeOH).

Step 2:N-(2-Benzyl-3-Mercaptopropionyl)-(S)-(4-Methyl-benzyl)-L-Cysteine(Isomer B)

Dissolve Isomer B (0.66 g, 1.6 mmole) in methanol (20 ml) under anitrogen atmosphere, cool to 0°-5°, add 1N sodium hydroxide (4.8 ml),stir the mixture at 0°-5° for 6 hours and then keep at that temperaturefor 18 hours. Concentrate the reaction mixture under nitrogen, dilutethe resultant oil with water (200 ml) and ethyl acetate (200 ml) andacidify to pH 2-4 with 1N hydrochloric acid. Dry (MgSO₄) the ethylacetate solution and concentrate in vacuo to give the title compound(Isomer B), a viscous oil (0.45 g), [α]_(D) ²⁶ =-56.3° (MeOH).

Step 3: N-(2-Benzyl-3-Mercaptopropionyl)-(S)-(4-Methylbenzyl)-L-Cysteine(Isomer A)

By a procedure similar to that of Step 2 react Isomer A (0.63 g) and 1Nsodium hydroxide (4.5 ml) to give the title compound (Isomer A), aviscous clear oil (0.165 g), [α]_(D) ²⁶ =-8.8° (MeOH).

In a similar manner, according to Example 1, Step 1, using theappropriate propionic acid, prepare:

N-[3-Acetylthio-2-(α-naphthylmethyl)propionyl]-S-(4-methylbenzyl)-L-cysteineethyl ester, Isomer A, m.p. 71°-74°, -40.6° (MeOH);

N-[3-Acetylthio-2-(α-naphthylmethyl)propionyl]-S-(4-methylbenzyl)-L-cysteineethyl ester, Isomer B, m.p. 88°-90°, [α]_(D) ²⁶ =-58.8° (MeOH);

N-[3-Acetylthio-2(β-naphthylmethyl)propionyl]-S-(4-methylbenzyl)-L-cysteineethyl ester, Isomer A, m.p. 74°-77°, [α]_(D) ²⁶ =-61.0° (MeOH);

N-[3-Acetylthio-2(β-naphthylmethyl)propionyl]-S-(4-methylbenzyl)-L-cysteineethyl ester, Isomer B, m.p. 86°-88°, [α]_(D) ²⁶ =-20.7° (MeOH);

N-[3-Acetylthio-2-(4-chlorobenzyl)propionyl]-S-benzyl-L-cysteine ethylester (Isomer A), m.p. 89°-90°;

N-[3-Acetylthio-2-(4-chlorobenzyl)propionyl]-S-benzyl-L-cysteine ethylester (Isomer B), m.p. 103°-4°; and

N-[3-Acetylthio-2-(4-chlorobenzyl)propionyl]-L-tryptophan methyl ester(Isomers A and B).

Using the procedure of Example 1, Step 2, treat the above 3-acetylthiocompounds to obtain the following 3-mercaptopropionyl compounds:

N-[2-(α-Naphthylmethyl)-3-mercaptopropionyl]-S-(4-methylbenzyl)-L-cysteineIsomer A, m.p. 70°-75°, [α]_(D) ²⁶ =+18.3° )MeOH);

N-[2-(α-Naphthylmethyl)-3-mercaptopropionyl]-S-(4-methylbenzyl)-L-cysteineIsomer B, m.p. 48°-55°, [α]_(D) ²⁶ =-102.3° (MeOH);

N-[2-(β-Naphthylmethyl)-3-mercaptopropionyl]-S-(4-methylbenzyl)-L-cysteine,Isomer A, a white foam, [α]_(D) ²⁶ =+9.9° (MeOH);

N-[2-(β-Naphthylmethyl)-3-mercaptopropionyl]-S-(4-methylbenzyl)-L-cysteine,Isomer B, a white foam, [α]_(D) ²⁶ =-50.1° (MeOH);

N-[2-(4-chlorobenzyl)-3-mercaptopropionyl]-S-benzyl-L-cysteine (IsomerA), [α]_(D) ²⁶ =-3.0° (EtOH, c=1);

N-[2-(4-chlorobenzyl)-3-mercaptopropionyl]-S-benzyl-L-cysteine (IsomerB), [α]_(D) ²⁶ =-48.7° (EtOH, c=1);

N-[2-(4-chlorobenzyl)-3-mercaptopropionyl]-L-tryptophan (Isomer A),[α]_(D) ²⁶ =+18.1° (EtOH, c=0.5); and

N-[2-(4-chlorobenzyl)-3-mercaptopropionyl]-L-tryptophan (Isomer B),[α]_(D) ²⁶ =-22.5° (EtOH, c=0.5).

EXAMPLE 2 N-(2-Benzyl-3-Mercaptopropionyl)-S-Benzyl-L-Cysteine (IsomersA & B) Step 1: N-(3-Acetylthio-2-Benzylpropionyl)-S-Benzyl-L-Cysteine,Ethyl Ester (Isomers A & B)

React S-benzyl-L-cysteine ethyl ester hydrochloride (1.38 g) and3-acetylthio-2-benzyl propionic acid (1.19 g) in a procedure similar tothat described in Example 1, Step 1 to give a yellow oil. Chromatographthe oil on silica gel (1.5L, 60-200 mesh) using CH₂ Cl₂ :ethyl acetate98:2 as eluant to give Isomer A, white solid (0.49 g), m.p. 83°-5°,[α]_(D) ²⁶ =-73.5° (MeOH); overlap Isomer A and B (0.66 g); and IsomerB, white solid, m.p. 72°-4°, [α]_(D) ²⁶ =-9.4° (MeOH).

Step 2

Using the procedure described in Example 1, Step 2, separately treat theIsomers of Step 1 above to obtain Isomers A and B of the title compound:Isomer A, a colorless oil, [α]_(D) ²⁶ =-2.1° (MeOH), and Isomer B, acolorless oil, [α]_(D) ²⁶ =-46.7° (MeOH).

EXAMPLE 3 N-(2-Benzyl-3-Mercaptopropionyl)-S-Benzyl-D-Cysteine (IsomersA AND B) Step 1: N-(3-Acetylthio-2-Benzylpropionyl)-S-Benzyl-D-Cysteine,Ethyl Ester (Isomers A and B)

React S-benzyl-D-cysteine ethyl ester hydrochloride (2.05 g) and3-acetylthio-2-benzylpropionic acid (1.77 g) in a manner similar to thatdescribed in Example 1, Step 1 to give a light amber oil. Place the oilon a column of silica gel (2L, 60-200 mesh) and elute with CH₂ Cl₂:ethyl acetate 98:2 to give Isomer A, white solid (0.70 g), m.p.84°-85°; [α]_(D) ²⁶ =+75.9° (MeOH); overlap Isomer A and Isomer B (0.85g); and Isomer B, white solid (0.33 g), [α]_(D) ²⁶ =+15.6° (MeOH).

Step 2

Using the procedure described in Example 1, Step 2, separately treat theIsomers of Step 1 above to obtain Isomers A and B of the title compound:Isomer A, a colorless oil, [α]_(D) ²⁶ =+38.2° (MeOH).

EXAMPLE 4N-(2-Benzyl-3-Mercaptopropionyl)-S-(4-Methoxybenzyl)-L-Cysteine (IsomersA AND B) Step 1:N-(3-Acetylthio-2-Benzylpropionyl)-S-(4-Methoxybenzyl)-L-Cysteine MethylEster (Isomers A and B)

React S-(4-methoxybenzyl)-L-cysteine methyl ester hydrochloride (1.85 g)and 3-acetylthio-2-benzylpropionic acid (1.95 g) in the manner describedin Example 1, Step 1 to give an amber oil. Chromatograph this oil on acolumn of silica gel (2L, 60-200 mesh) and elute with ethylacetate:hexane 5:20 to give Isomer A, a clear oil (0.63 g), [α]_(D) ²⁶=-66.5° (MeOH), overlap Isomer A and Isomer B (0.28 g); and Isomer B, aclear oil (0.67 g), [α]_(D) ²⁶ =+3.0° (MeOH).

Step 2

Using the procedure described in Example 1, Step 2 separately treat theisomers of Step 1 above to obtain Isomers A and B of the title compound:Isomer A, a viscous oil, [α]_(D) ²⁶ =-19.3° (MeOH); and Isomer B, aviscous oil, [α]_(D) ²⁶ =-44.2° (MeOH).

EXAMPLE 5N-(2-Benzyl-3-Mercaptopropionyl)-S-(3,4-Dimethylbenzyl)-L-Cysteine(Isomers A and B) Step 1:N-(3-Acetylthio-2-Benzylpropionyl)-S-(3,4-Dimethylbenzyl)-L-CysteineEthyl Ester (Isomers A and B)

React S-(3,4-dimethylbenzyl)-L-cysteine ethyl ester hydrochloride (2.20g) and 3-acetylthio-2-benzylpropionic acid (1.74 g) in the mannerdescribed in Example 1, Step 1 to give an amber oil. Place the oil on acolumn of silica gel (1L) and elute with ethyl acetate:hexane 25:170(4L) and then methanol:hexane 25:170 to give a light orange oily solid.Repeat the chromatography to give Isomer A, a white solid (0.52 g), m.p.89.5°-92.5°, [α]_(D) ²⁶ =-71.1° (MeOH) and Isomer B, a white solid (0.60g), m.p. 51°-55°, [α]_(D) ²⁶ =-8.4° (MeOH).

Step 2

Using the procedure described in Example 1, Step 2, separately treat theIsomers of Step 1 above, to obtain Isomers A and B of the titlecompound: Isomer A, a clear viscous oil, [α]_(D) ²⁶ =-18.0° (MeOH); andIsomer B, a clear viscous oil, [α]_(D) ²⁶ =-56.5° (MeOH).

EXAMPLE 6 N-(2-Benzyl-3-Mercaptopropionyl)-S-Phenethyl-L-Cysteine(Isomers A and B) Step 1:N-(3-Acetylthio-2-Benzylpropionyl)-S-Phenethyl-L-Cysteine Ethyl Ester,(Isomers A and B)

React S-phenethyl-L-cysteine ethyl ester hydrochloride (2.85 g) and3-acetylthio-2-benzylpropionic acid, (2.38 g) in a manner similar tothat described in Example 1, Step 1 to give an amber oil. Chromatographthis oil on Prep 500 (2 silica gel cartridges) and elute with CH₂ Cl₂(4L) and then CH₂ Cl₂ :ethyl acetate 100:2 to give Isomer A, a whitesolid (1.32 g), m.p. 63°-64°[α]_(D) ²⁶ =-51.2° (MeOH); overlap Isomer Aand Isomer B (0.63 g); and Isomer B, white solid (1.14 g), m.p. 84°-86°,[α]_(D) ²⁶ =+5.3° (MeOH).

Step 2

Using the procedure described in Example 1, Step 2, separately treatIsomers A and B of Step 1 above to obtain Isomers A and B of the titlecompound. Isomer A, a colorless oil, [α]_(D) ²⁶ =+4.8° (MeOH); andIsomer B, a colorless oil, [α]_(D) ²⁶ =-39.7° (MeOH).

EXAMPLE 7 N-(2-Benzyl-3-Mercaptopropionyl)-S-(t-Butyl)-L-Cysteine(Isomers A and B) Step 1:N-(3-Acetylthio-2-Benzylpropionyl)-S-(t-Butyl)-L-Cysteine Methyl Ester(Isomers A and B)

React S-(t-butyl)-L-cysteine methyl ester (2.32 g) and3-acetylthio-2-benzylpropionic acid (3.22 g) in the manner described inExample 1, Step 1 to give an orange solid. Chromatograph this solid on acolumn of silica gel (2L, 60-200 mesh) and elute with ethylacetate:hexane 3:17 to give Isomer A, a clear oil (1.09 g), [α]_(D) ²⁶=-44.9° (MeOH); overlap Isomer A and Isomer B (0.52 g); and Isomer B, aclear oil (0.75 g), [α]_(D) ²⁶ =+8.3° (MeOH).

Step 2

Using the procedure described in Example 1, Step 2, separately treat theIsomers above to obtain Isomers A and B of the title compound; Isomer A,a clear viscous oil, [α]_(D) ²⁶ =+0.4° (MeOH), and Isomer B, a whitesolid, m.p. 68°-75°, [α]_(D) ²⁶ =-32.3° (MeOH).

EXAMPLE 8 N-(2-Benzyl-3-Mercaptopropionyl)-L-Ethionine (Isomers A and B)Step 1: N-(3-Acetylthio-2-Benzylpropionyl)-L-Ethionine Ethyl Ester(Isomers A and B)

React L-ethionine ethyl ester (3.51 g) and3-acetylthio-2-benzylpropionic acid (4.37 g) in a manner similar to thatdescribed in Example 1, Step 1 to give a yellow residue. Chromatographthe yellow residue on the Waters Prep 500 (2 silica gel cartridges) andelute with ethyl acetate:hexane 2:18 (16L), then ethyl acetate:hexane3:17. Repeat chromatography of the fractions using ethyl acetate:hexaneas eluant to give Isomer A, a white solid (0.89 g), m.p. 84°-90°,[α]_(D) ²⁶ =-60.6° (MeOH) and Isomer B (0.82 g), m.p. 79°-84°, [α]_(D)²⁶ =-0.3° (MeOH).

Step 2

Using the procedure described in Example 1, Step 2, separately treat theIsomers of Step 1 above to obtain Isomers A and B of the title compound;Isomer A, a milky viscous oil, [α]_(D) ²⁶ =-41.8° (MeOH); and Isomer B,a milky viscous oil, [α]_(D) ²⁶ =-66.0° (MeOH).

EXAMPLE 9 N-(2-Benzyl-3-Mercaptopropionyl)-O-Benzyl-L-Tyrosine (IsomersA and B) Step 1: N-(3-Acetylthio-2-benzylpropionyl)-O-benzyl-L-tyrosineMethyl Ester (Isomers A and B)

React O-benzyl-L-tyrosine methyl ester hydrochloride (2.,7 g) and3-acetylthio-2-benzylpropionic acid (2.05 g) in a manner similar to thatdescribed in Example 1, Step 1 to give a yellow-orange oil.Chromatograph this oil on a column of silica gel (2.5L) and elute withCH₂ Cl₂ :ethyl acetate 98:2 to give Isomer A, a white solid (0.84 g)m.p. 108°-109°; [α]_(D) ²⁶ =-39.8° (MeOH); overlap Isomer A and Isomer B(0.80 g); and Isomer B, white solid (0.45 g), m.p. 92°-93°, [α]_(D) ²⁶=+19.2° (MeOH).

Step 2

Using the procedure described in Example 1, Step 2, separately treat theIsomers of Step 1 above to obtain Isomers A and B of the title compound:Isomer A, an off-white solid, [α]_(D) ²⁶ =+4.8° (MeOH); and Isomer B, aviscous colorless oil [α]_(D) ²⁶ =+2.4° (MeOH).

EXAMPLE 10 N-(2-Benzyl-3-Mercaptopropionyl)-(S)-Bishomophenyl Alanine(Isomers A and B) Step 1:N-(3-Acetylthio-2-Benzylpropionyl)-(S)-Bishomophenylalanine t-ButylEster (Isomers A and B)

React (S)-bishomophenylalanine t-butyl ester hydrochloride (2.49 g) and3-acetylthio-2-benzylpropionic acid (2.39 g) in the manner described inExample 1, Step 1 to give a yellow oil. Chromatograph this oil on WatersPrep 500 (2 silica gel cartridges) and elute with CH₂ Cl₂ (4L) and thenCH₂ Cl₂ :ethyl acetate 100:2 to give Isomer A, a colorless oil (0.99 g),[α]_(D) ²⁶ =-54.7° (MeOH), overlap Isomer A and Isomer B (0.62 g); andIsomer B, a colorless oil (0.79 g), [α]_(D) ²⁶ =+5.1° (MeOH).

Step 2: N-(3-Acetylthio-2-Benzylpropionyl)-(S)-Bishomophenylalanine(Isomer A and B)

To Isomer A of the product of Step 1 (0.97 g, 0.21 mmole) in CH₂ Cl₂ (10ml) at 0°-5°, add dropwise trifluoroacetic acid (10 ml). Warm theresulting mixture to room temperature, stir for 18 hours, andconcentrate in vacuo. Dissolve the residue in CH₂ Cl₂ (10 ml) andconcentrate in vacuo. Treat the residue with diethyl ether (10 ml) andconcentrate in vacuo to give Isomer A of the title compound, a lightamber oil (0.87 g), [α]_(D) ²⁶ =-43.0° (MeOH).

By this same method, convert Isomer B of Step 1 toN-(3-acetylthio-2-benzylpropionyl)-(S)-bishomophenylalanine Isomer B, alight amber oil, [α]_(D) ²⁶ =+19.6° (MeOH).

Step 3: N-(2-Benzyl-3-Mercaptopropionyl)-(S)-Bishomophenylalanine(Isomers A and B)

Dissolve Isomer A of Step 2 in methanol (15 ml) at 0°-5° under anitrogen atmosphere and treat with 1N sodium hydroxide (6.3 ml). Treatthe resulting mixture as described in Example 1, Step 2, to give IsomerA of the title compound, a pale yellow viscous oil (0.69 g), [α]_(D) ²⁶=-25.4° (MeOH).

By this same method, convert Isomer B of Step 2 toN-(2-benzyl-3-mercaptopropionyl)-(S)-bishomophenylalanine Isomer B, apale yellow viscous oil, [α]_(D) ²⁶ =-50.0° (MeOH).

In a similar manner, substitute (S)-(4-methylbenzyl)-L-cysteine t-butylester (Preparation 6) for (S)-bishomophenylalanine in Example 10, Step 1to obtain

N-(3-acetylthio-2-benzylpropionyl)-(S)-(4-methylbenzyl)-L-cysteinet-butyl ester, Isomer A, [α]_(D) ²⁶ =-82.8° (MeOH); and

N-(3-acetylthio-2-benzylpropionyl)-(S)-(4-methylbenzyl)-L-cysteinet-butyl ester, Isomer B, [α]_(D) ²⁶ =-23.5° (MeOH).

Treat the above esters in a manner similar to that described in Example10, Step 2 to obtain

N-(3-acetylthio-2-benzylpropionyl)-(S)-(4-methylbenzyl)-L-cysteine,Isomer A, [α]_(D) ²⁶ =-58.8° (MeOH); and

N-(3-acetylthio-2-benzylpropionyl)-(S)-(4-methylbenzyl)-L-cysteine,Isomer B, [α]_(D) ²⁶ =-6.6° (MeOH).

EXAMPLE 11 N-[3-Mercapto-2(R,S)-Benzylpropionyl]-L-Methionine Step 1:N-[3-Acetylthio-2(R,S)-Benzylpropionyl]-L-Methionine Methyl Ester

Add methionine methyl ester (2.00 g, 1.23 mmole) to3-acetylthio-2-benzylpropionic acid (3.01 g, 1.26 mmole), DEC (2.34 g,1.22 mmole), HOBT (1.88 g, 1.23 mmole) and NMM (2.34 g, 2.31 mmole) inDMF (12 ml), and treat the resulting mixture as described in Example 1,Step 1 to give an amber oil (4.36 g). Chromatograph the oil on a columnof silica gel (1L 60-200 mesh) elute with CH₂ Cl₂ (1L) and then CH₂ Cl₂:ethyl acetate 99:1 to give the title compound, a clear oil (2.61 g),[α]_(D) ²⁶ =-38.9° (MeOH).

Step 2: N-[3-Mercapto-2(R,S)-Benzylpropionyl]-L-Methionine

Dissolve the product of Step 1 in methanol (20 ml) and treat with 1NNaOH (20.4 ml) as described in Example 1, Step 2 to give the titlecompound, a white solid, m.p. 132°-5°, [α]_(D) ²⁶ =-34.6° (MeOH).

Using the method of Example 11, Step 1, otherN-[3-(R,S)-Acetylthio-2-benzylpropionyl]amino acid esters are prepared:

N-[3-Acetylthio-2(R,S)-benzylpropionyl]-S-methyl-L-cysteine ethyl ester,a clear oil, [α]_(D) ²⁶ =-25.9° (MeOH);

N-[3-Acetylthio-2(R,S)-benzylpropionyl]-S-trityl-L-cysteine methylester, an amber oil, [α]_(D) ²⁶ =+5.9° (MeOH); and

N-[3-Acetylthio-2(R,S)-benzylpropionyl]-(S)-tryptophan methyl ester, acolorless oil, [α]_(D) ²⁶ =-14.7° (MeOH).

Similarly, from 3-benzoylthio-2(S)-benzylpropionic acid, prepareN-(3-benzoylthio-2(S)-benzylpropionyl)-S-methionine methyl ester, awhite solid, m.p. 78°-80° C., [α]_(D) ²⁶ =-67.9° (EtOH, c=1).

Using the procedure of Example 11, Step 2, convert the above acetylthioand benzoylthio compounds to the following 3-mercaptopropionylcompounds:

N-[2(R,S)-Benzyl-3-mercaptopropionyl]-S-methyl-L-cysteine, a clearviscous oil, [α]_(D) ²⁶ =-31.1° (MeOH);

N-[2(R,S)-Benzyl-3-mercaptopropionyl)-S-trityl-L-cysteine, a whitesolid, [α]_(D) ²⁶ =+10.5° (MeOH);

N-[2(R,S)-Benzyl-3-mercaptopropionyl)-(S)-tryptophan, a white foam, m.p.68°-69°, [α]_(D) ²⁶ =-0.5° (MeOH); and

N-[2(S)-Benzyl-3-mercaptopropionyl)-S-methionine, a white solid, m.p.88°-89°.

EXAMPLE 12N-[2-(4-Phenylbenzyl)-3-Mercaptopropionyl)-S-(4-Methylbenzyl)-L-Cysteine(S,R and R,R Diastereomers) Step 1:3-Acetylthio-2-(4-Phenylbenzyl)Propionyl Chloride

To 3-acetylthio-2-(4-phenylbenzyl)propionic acid (3.39 g, 10.8 mmole) intoluene (25 ml) add 1% DMF in toluene (2 drops) and thionyl chloride(1.2 ml, 1.65 g, 13.8 mmoles) and stir the resulting solution at roomtemperature for 18 hours. Concentrate the reaction mixture in vacuo,dissolve the residue in toluene (100 ml) and concentrate the solution invacuo to give the title compound, a light brown oil (3.37 g).

Step 2:N-[3-Acetylthio-2-(4-Phenylbenzyl)Propionyl]-S-4-Methylbenzyl)-L-Cysteine(S,R and R,R diastereomers)

Add the acid chloride (3.37 g) from Step 1 in acetonitrile (25 ml)dropwise to S-(4-methylbenzyl)-L-cysteine hydrochloride (2.62 g, 10mmol) in acetonitrile (30 ml), water (15 ml) and triethylamine (2.8 ml),and stir the resulting mixture at room temperature for 4 hours.Concentrate the reaction mixture in vacuo and partition the residuebetween ethyl acetate (700 ml) and water (2×200 ml) and then saturatedsodium chloride solution (100 ml). Dry the ethyl acetate solution(MgSO₄) and concentrate in vacuo to give a brown solid. Chromatographthis solid on a column of silica gel (2L, 60-200 mesh) and elute withCH₂ Cl₂ :methanol:glacial acetic acid (97.5:2.5:0.25) to give a whitefoam (2.45 g). Chromatograph this white foam on a column of silica gel(1.2L, 60-200 mesh) and elute with CH₂ Cl₂ methanol:glacial acetic acid(97.5:2.5:0.25) to give the S,R-diastereomer of title compound, a whitesolid (1.04 g), m.p. 123°-125°, [α]_(D) ²⁶ =-45.5° (EtOH) and theR,R-diastereomer of the title compound, a white solid (0.86 g), m.p.131°-135°, [α]_(D) ²⁶ =-7.1° (MeOH).

Using a similar procedure, prepare the diastereomers of the following:

N-[3-acetylthio-2-benzylpropionyl]-S-(4-methylbenzyl)-L-cysteine: theS,R-diastereomer, a colorless oil, [α]_(D) ²⁶ =-23.0° (MeOH) and theR,R-diastereomer, a yellow oil, [α]_(D) ²⁶ =-1.3° (MeOH);

N-(2-acetythiomethyl-3-(4-methylphenylpropionyl)-S-methionine: theS,S-diastereomer, a white solid, m.p. 90°-93°, [α]_(D) ²⁶ =-32.2° (EtOH,c=1) and the R,S-diastereomer, a white solid, [α]_(D) ²⁶ =+13.0° (EtOH,c=1);

N-(2-acetylthiomethyl-3-phenylpropionyl)-S-methionine: theS,S-diastereomer, a white solid, m.p. 95°-98°, [α]_(D) ²⁶ =-36.6° (EtOH,c=1).

Step 3:N-[2-(4-Phenylbenzyl)-3-Mercaptopropionyl]-S-(4-Methylbenzyl)-L-Cysteine(S,R and R,R-diastereomers)

Dissolve the S,R-diastereomer of Step 2 in methanol saturated withammonia (50 ml) at 0°-5° under a nitrogen atmosphere. After 35 minutes,bubble nitrogen through the reaction mixture. Dilute the reactionmixture with water and acidify to pH 2-4 with 1N hydrochloric acid.Extract the acidic solution with ethyl acetate, dry the organic layer(MgSO₄) and concentrate in vacuo to give the S,R-diastereomer of thetitle compound, a white solid (0.73 g). Purify the S,R-diastereomer byflash chromatography on silica gel (Baker flash silica gel, 40 μM) (25g) eluting with CHCl₂ :MeOH:gl.AcOH, 97.5:2.5:0.25, to obtain a whitesolid (0.549 g), [α]_(D) ²⁶ =-2.2° (MeOH).

In a similar fashion, prepare the R,R-diastereomer of the titlecompound, a white solid, [α]_(D) ²⁶ =-62.2° (MeOH).

EXAMPLE 13 N-[2-Benzyl-3-Mercaptopropionyl]-L-Methionine Amide Step 1:N-(3-Acetylthio-2-Benzylpropionyl)-L-Methionine Amide

In similar fashion to that described in Example 12, Step 2, convertL-methionine amide to N-(3-acetylthio-2-benzylpropionyl)-L-methionineamide. Recrystallize from hexane:CH₂ Cl₂ to obtain a solid, m.p.101°-3°. Chromatograph on silica gel with 4% methanol/CH₂ Cl₂ to obtainthe S,S-diastereomer, m.p. 149°-51°, [α]_(D) ²⁶ =87.5° (CHCl₃) and theR,S-diastereomer, m.p. 119°-21°, [α]_(D) ₂₆ = +5.0° (CHCl₃).

Step 2 N-(2-Benzyl-3-Mercaptopropionyl)-L-Methionine Amide

Treat each of the 3-acetylthio diastereomers of Step 1 with NH₃ /MeOHfor 4 hours as in Example 12, Step 3, to give the S,S-diastereomer ofthe title compound, [α]_(D) ²⁶ =-67.8° (EtOH, c=1), and theR,S-diastereomer of the title compound, m.p. 153°-6°, [α]_(D) ²⁶ =+9.7°(EtOH, c=1).

In a similar manner to that described in Example 13, Step 1, substitutethe appropriate acetylthio compounds and amides and separate bychromatography to obtain:

N-(3-Acetylthio-2-benzylpropionyl)-S-(4-methylbenzyl)-L-cysteine amide,Isomer A, [α]_(D) ²⁶ =-38.2° (MeOH);

N-(3-Acetylthio-2-benzylpropionyl)-S-(4-methylbenzyl)-L-cysteine amide,Isomer B, [α]_(D) ²⁶ =-1.6° (MeOH);

N-[3-Acetylthio-2-(4-chlorobenzylpropionyl)]-L-methionine amide, IsomerA;

N-[3-Acetylthio-2-(4-chlorobenzylpropionyl)]-L-methionine amide, IsomerB, m.p. 166°-9°;

N-(3-Acetylthio-2(S)-phenylpropionyl)-S-3-(2-thienyl)alanine amide, awhite solid, TLC R_(f) =0.35 (1% EtOH/EtOAc); and

N-(3-Acetylthio-2(R)-phenylpropionyl)-S-3-(2-thienyl)alanine amide, TLCR_(f) =0.30 (1% EtOH/EtOAc).

Treat the amides obtained above in a manner similar to that described inExample 13, Step 2 to obtain:

N-[2-(4-Chlorobenzyl)-3-mercaptopropionyl]-L-methionine amide, Isomer A,m.p. 194°, [α]_(D) ²⁶ =+1.2° (MeOH);

N-[2-(4-Chlorobenzyl)-3-mercaptopropionyl]-L-methionine amide, Isomer B,[α]_(D) ²⁶ =-65.2° (MeOH);

N-(2-Benzyl-3-mercaptopropionyl)-S-(4-methylbenzyl)-L-cysteine amide,Isomer A, m.p. 130°-2°, [α]_(D) ²⁶ =-4.2° (MeOH);

N-(2-Benzyl-3-mercaptopropionyl)-S-(4-methylbenzyl)-L-cysteine amide,Isomer B, [α]_(D) ²⁶ =-29.0° (MeOH);

N-(2(S)-Benzyl-3-mercaptopropionyl)-S-3-(2-thienyl)alanine amide, awhite solid, TLC R_(f) =0.40 (4% MeOH/CH₂ Cl₂); and

N-(2(R)-Benzyl-3-mercaptopropionyl)-S-3-(2-thienyl)alanine amide, TLCR_(f) =0.35 (4% MeOH/CH₂ Cl₂).

EXAMPLE 14 N-(3-Benzoylthio-2-Benzylpropionyl)-L-Methionine Amide

Prepare the R and S enantiomers of 3-benzoylthio-2-benzylpropionic acidaccording to the procedure described in U.S. Pat. No. 4,329,495, hereinincorporated by reference.

In a manner similar to that described in Example 1, Step 1, condenseeach acid separately with L-methionine amide to obtain

N-(3(S)-benzoylthio-2-benzylpropionyl)-L-methionine amide, m.p.178°-180°, [α]_(D) ²⁶ =-97.6° (MeOH); and

N-(3(R)-benzoylthio-2-benzylpropionyl)-L-methionine amide, m.p. 145°-9°,[α]_(D) ²⁶ =+32.9° (CHCl₃).

EXAMPLE 15 N-[2-Benzyl-3-Mercaptopropionyl]-L-Aspartic Acid β-BenzylEster

In similar fashion to that described in Example 12, Step 2, convertL-aspartic acid β-benzyl ester toN-(3-acetylthio-2-benzylpropionyl)-L-aspartic acid, β-benzyl ester.

Treat with NH₃ /MeOH as in Example 12, Step 3 to give the titlecompound, [α]_(D) ²⁶ =-5.7° (EtOH, c=0.5).

EXAMPLE 16 N-[N-(2-Benzyl-3-Mercaptopropionyl)-L-Phenylalanyl]-L-Alanine

Using the procedure of Example 1, convert L-phenylalanyl-L-alaninebenzyl ester hydrochloride toN-(N-(3-acetylthio-2-benzylpropionyl)-L-phenylalanyl]-L-alanine, benzylester.

Treat with NaOH as in Example 1, Step 2 to give the crude titlecompound. React the product (0.62 g) with zinc powder (0.5 g) and 5N HCl(10ml) in 20ml MeOH for 1 hour. Concentrate the mixture, extract withCH₂ Cl₂, dry, and remove the solvent to give the title compound, [α]_(D)²⁶ =-23.1° (EtOH, c=0.5).

Use the same procedure to prepareN-(N-(2-benzyl-3-mercaptopropionyl)-L-phenylalanyl-L-leucine, [α]_(D) ²⁶=-20.8° (EtOH, c=0.5).

EXAMPLE 17 N-[N-(2-Benzyl-3-Mercaptopropionyl)-L-Alanyl]-L-Proline Step1: N-[N-(3-Acetylthio-2-Benzylpropionyl)-L-Alanyl]-L-Proline

Using the procedure of Example 12, Step 2, convert L-alanyl-L-proline toN-[N-(3-acetylthio-2-benzylpropionyl)-L-alanyl]-L-proline, a white foam,[α]_(D) ²⁶ =-81.8° (MeOH).

Step 2: N-[N-(2-Benzyl-3-Mercaptopropionyl)-L-Alanyl]-L-Proline

Treat the product from Step 1 with methanol saturated with ammonia asdescribed in Example 12, Step 3 (before chromatography). Treat theresultant residue with zinc powder as described in Example 17.Chromatograph the product on flash grade silica gel using CH₂ Cl₂:MeOH:NH₄ OH (97.5:2.5:0.25) to give the title compound, [α]_(D) ²⁶=-118.2° (MeOH).

EXAMPLE 18 N-(2(R,S)-Acetylthiomethyl-3-Phenylpropionyl)-S-MethioninePiperidine Amide Step 1

Combine N-(t-butoxycarbonyl)-S-methionine (1.50 g, 6.0 mmol), piperidine(0.51 g, 6.0 mmol) and HOBT (0.91 g, 6.0 mmol) in DMF (40 ml). Add DEC(1.15 g, 6.0 mmol). Stir 5 hours, concentrate in vacuo, and partitionbetween EtOAc and H₂ O. Wash with aqueous NaHCO₃, dry, and concentrateto obtain the piperidine amide as a colorless oil (TLC single spot R_(f)=0.6 in 5% MeOH/CH₂ Cl₂).

Step 2: Treat the amide of Step 1 with 6M HCl/dioxane (25 ml). After 0.5hr., concentrate in vacuo to obtain the amine hydrochloride as a foam.Step 3

Treat the above amide hydrochloride of Step 2 with triethylamine (1.35g, 15 mmol) in CH₃ CN:H₂ O 2:1 (60 ml). Add2-acetylthiomethyl-3-phenylpropionyl chloride (1.37 g, 5.4 mmol), stir 1hr., add 1N HCL (15 ml), and extract with EtOAc. Dry and concentrate toobtain a brown oil. Chromatograph on silica gel (2% MeOH/CH₂ Cl₂ aseluent) to obtain the product, a 3:2 mixture of S,S-andR,S-diastereomers, as a yellow oil, [α]_(D) ²⁶ =-31.7° (EtOH, c=1).

Using a similar procedure, prepare the following mixtures ofdiastereomers:

N-(2(R,S)-acetylthiomethyl-3-phenylpropionyl)-S-methionine4-methylbenzyl amide, a 1:1 mixture of diastereomers, a colorless foam,[α]_(D) ²⁶ =-18.6° (EtOH, c=1);

N-(2(R,S)-acetylthiomethyl-3-phenylpropionyl)-S-methionine2-hydroxyethyl amide, a 3:2 mixture of the S,S- and R,S-diastereomers,[α]_(D) ²⁶ =-31.1° (EtOH, c=1); and

N-(2(R,S)-acetylthiomethyl-3-phenylpropionyl)-S-methionine morpholineamide, a 1:1 mixture of diastereomers, a yellow oil, [α]_(D) ²⁶ =-30.0°(EtOH, c=1).

Using a similar procedure, prepareN-(2(R,S)-acetylthiomethyl-3-phenylpropionyl)-S-methionine pyrrolidineamide. Chromatograph on silica gel (elute with diethylether) to obtainthe S,S-diastereomer as a yellow solid, [α]_(D) ²⁶ =-49.4° (EtOH, c=1)and the R,S-diastereomer as a white solid, [α]_(D) ²⁶ =-6.1° (EtOH,c=1).

Again, using a procedure similar to Example 18, prepareN-(2(S)-acetylthiomethyl-3-phenylpropionyl)-S-methionine2-dimethylaminoethyl amide, a beige solid, [α]_(D) ²⁶ =-58.3° (EtOH,c=1). Dissolve in CH₂ Cl₂, add HCl/Et₂ O and concentrate to obtain thehydrochloride salt as a yellow gum.

EXAMPLE 19 N-(2(S)-Acetylthiomethyl-3-Phenylpropionyl)-S-Methionine2-(2-Chloroethoxy)Ethyl Ester Step 1

Combine N-(t-butoxycarbonyl)-S-methionine (2.0 g, 8.0 mmol),2-(2-chloroethoxy)ethanol (0.99 g, 8.0 mmol) and 4-dimethylaminopyridinein CH₂ Cl₂ (35 ml). Add DEC (1.54 g, 8.0 mmol). Stir 1 hour,concentrate, and partition between Et₂ O and 1N HCl. Dry and concentrateto obtain the ester as an oil.

Step 2

Treat the ester of Step 1 with 6M HCl/dioxane (25 ml). After 35 min.,concentrate and triturate the residue with Et₂ O to obtain the aminehydrochloride as a solid.

Step 3

Treat the hydrochloride of Step 2 (1.30 g, 4.5 mmol) with triethylamine(0.90 g, 8.9 mmol) in CH₃ CN:H₂ O 2:1 (60 ml). Add2-acetylthiomethyl-3-phenylpropionyl chloride (1.14 g, 4.5 mmol), stir 1hour, add 1N HCl (13 ml), and extract with EtOAc. Dry and concentrate.Chromatograph on silica gel (Et₂ O/hexane 6:4 as eluent) to obtain thetitle compound as a yellow solid, m.p. 58°-63°, [α]_(D) ²⁶ =-44.7°(EtOH, c=1).

In a similar manner, prepare mixtures of esters and separate bychromatography to obtain:

N-(2(S)-acetylthiomethyl-3-(4-methyphenyl)-propionyl)-S-methionine2-(2-chloroethoxy)ethyl ester, a yellow solid, [α]_(D) ²⁶ =-47.7° (EtOH,c=1);

N-(2(R)-acetylthiomethyl-3-(4-methyphenyl)-propionyl)-S-methionine2-(2-chloroethoxy)ethyl ester, a yellow solid, [α]_(D) ²⁶ =-2.5° (EtOH,c=1);

N-(2(S)-acetylthiomethyl-3-(2-methylphenyl)propionyl)-S-methionine2-(2-chloroethoxy) ethyl ester, a yellow solid, [α]_(D) ²⁶ =-46.4°(EtOH, c=1); and

N-(2(R)-acetylthiomethyl-3-(2-methylphenyl)-propionyl)-S-methionine2-(2-chloroethoxy) ethyl ester, a yellow solid, [α]_(D) ²⁶ =-17.7°(EtOH, c=1).

EXAMPLE 20 N-(2(R,S)-Acetylthiomethyl-3-Phenylpropionyl-S-MethionineDiethylaminocarbonylmethyl Ester Step 1

Combine N-(t-butoxycarbonyl)-S-methionine (1.50 g, 6.0 mmol),N,N-diethylbromoacetamide (1.16 g, 6.0 mmol) and Cs₂ CO₃ (0.97 g, 3.0mmol) in DMF (40 ml), stir 18 hours, and partition between EtOAc and H₂O. Wash with 1N NaHC₃, dry and concentrate to obtain the ester as anoil.

Step 2

Treat the ester as in Example 19, Steps 2 and 3, to obtain the titlecompound as a 3:2 mixture of S,S- and R,S-diastereomers, yellow oil,[α]_(D) ²⁶ =-30.4° (EtOH, c=1).

EXAMPLE 21 N-(2(S)-Benzoylthiomethyl-3-Phenylpropionyl)-S-Methionine2-(2-Chloroethoxy)Ethyl Ester

Combine the product of Example 19, Step 2 (1.00 g, 3.4 mmol) with2(S)-benzoylthiomethyl-3-phenylpropionic acid (see U.S. Pat. No.4,329,495) (1.03 g, 3.4 mmol), HOBT (0.52 g, 3.4 mmol) and triethylamine(0.86 g, 8.6 mmol) in DMF (35 ml). Add DEC (0.65 g, 3.4 mmol) and stir18 hours. Concentrate and partition between EtOAc and H₂ O. Wash with 1NNaHCO₃, dry and concentrate. Chromatograph the resultant oil on silicagel (Et₂ O:hexane 3:1 as eluant) to obtain the title compound, a whitesolid, [α]_(D) ²⁶ =-50.6° (EtOH, c=1).

The R,S-diastereomer of the title compound may be prepared in a similarmanner.

In a similar manner, prepare the following esters ofN-(2(S)-benzoylthiomethyl-3-phenylpropionyl)-S-methionine:

2-(ethoxy)ethyl ester, a yellow solid, [α]_(D) ²⁶ =-49.9° (EtOH, c=1);

2-(2-methoxyethoxy)ethyl ester, a white solid, [α]_(D) ²⁶ =-48.9° (EtOH,c=1);

2,2-dimethyl-1,3-dioxolan-4-yl or methyl ester, a white solid, m.p.76°-8°, [α]_(D) ²⁶ =-58.6° (MeOH, c=1).

The last compound may be converted to the 2,3-dihydroxypropyl ester byhydrolysis with AGW-50 resin (H⁺ form) in CH₃ CN:H₂ O 2:1 for 20 hoursand subsequent silica gel chromatography. The product ester is a whitesolid, [α]_(D) ²⁶ =-57.4° (MeOH, c=1).

EXAMPLE 22N-(3-Phenyl-2(S)-(4-Pyridylcarbonylthiomethyl)Propionyl)-S-MethionineAmide andN-(3-Phenyl-2(R)-(4-Pyridylcarbonylthiomethyl)Propionyl)-S-MethionineAmide

To N-(2(R,S)-mercaptomethyl-3-phenylpropionyl)-S-methionine amide (0.32g, 1.0 mmol) and triethylamine (0.19 g, 1.9 mmol) in CH₂ Cl₂ (30 ml),add isonicotinoyl chloride hydrochloride (0.19 g, 1.1 mmol). After 3hours, concentrate and chromatograph on silica (6% MeOH/CH₂ Cl₂ aseluent) to obtain:

the S,S-diastereomer, a yellow solid, m.p. 157°-60°, [α_(D) ²⁶ =-68.3°(EtOH, c=1); and

the S,R-diastereomer, a white solid, [α]_(D) ²⁶ =+27.6° (EtOH, c=1).

EXAMPLE 23 N-(2(R,S)-Mercaptomethyl-3-Phenylpropionyl)-S-Tryptophan4-Methylpiperazine Amide Hydrochloride andN-(2(R,S)-Mercaptomethyl-3-Phenylpropionyl)-S-Tryptophan2-(4-Pyridyl)Ethyl Amide Hydrochloride Step 1

Using the procedure of Example 18, prepareN(2(R,S)-acetythiomethyl-3-phenylpropionyl)-S-tryptophan4-methylpiperazine amide and convert to the hydrochloride, a brown foam,mass spectrum M⁺ =506.

Similarly, prepare the hydrochloride of the 2-(4-pyridyl)ethyl amide, anoff-white foam, mass spectrum M⁺ =528.

Step 2

Using the procedure of Example 12, Step 3, convert the products of Step1 to the title compounds:

the 4-methylpiperazine amide hydrochloride, an off-white foam, massspectrum M⁺ =486; and

the 2-(4-pyridyl)ethyl amide hydrochloride, a beige foam, mass spectrumM⁺ =464.

EXAMPLE 24 N-(2-Benzoylthiomethyl-3-Phenylpropionyl)-Methionine4-Methylpiperazine Amide Hydrochloride (S,S-and R,S-diastereomers)

Using procedures similar to Example 18, Step 1 and Example 1, Step 1,prepare the S,S-diastereomer of the title compound, a white solid, m.p.157°-8°. Similarly, prepare the R,S diastereomer, a white solid, m.p.114°-6°.

In a similar manner prepare the diastereomeric mixtureN-[(2(R,S)-benzoylthiomethyl-3-phenylpropionyl]-S-methionine2-(4-pyridyl)ethyl amide, a white solid, m.p. 108°-110°.

EXAMPLE 25 N-(2(S)-Benzoylthiomethyl-3-Phenylpropionyl)-S-MethionineSulfoxide 2-(2-Chloroethoxy)Ethyl Ester

To the title compound of Example 21 (1.00 g, 1.86 mmol) in MeOH (25 ml)add 30% H₂ O₂ (0.22 g, 4 mmol), stir 45 min. and concentrate. Trituratethe residue with Et₂ O:hexane 3:1 to obtain the title compound, a whitesolid, as a 1:1 mixture of sulfoxide diastereomers, NMR (CDCl₃) δ2.42,2.45.

In a similar manner, prepare the sulfoxide of the R,S-diastereomer ofthe title compound, a colorless oil, [α]_(D) ²⁶ =-2.8° (EtOH, c=1).

Also in a similar manner prepare the sulfoxide ofN-(2(S)-acetythiomethyl-3-phenylpropionyl)-S-methionine amide, a beigesolid, [α]_(D) ²⁶ =-50.6° (EtOH, c=1).

EXAMPLE 26 N-(2(S)-Mercaptomethyl-3-Phenylpropionyl)-S-MethionineSulfoxide

Treat the title compound of Example 25 (the S,S-diastereomer) (0.46 g,1.0 mmol) with MeOH (8 ml) and 1N NaOH (4 ml). Stir 18 hr. under N₂,concentrate and add 1N HCl (4 ml). Extract with EtOAc, dry, concentrateand chromatograph on silica (CH₂ Cl₂ :MeOH:HOAc 90:9:1 as eluent) toobtain the title compound as a white solid, NMR (DMSO) δ2.45, 2.46.

We claim:
 1. A method of treating congestive heart failure comprisingadministering an effective amount of a compound having the structuralformula ##STR61## wherein R^(1a) is Y--C₆ H₄ --, Y--C₆ H₄ S--, Y--C₆ H₄O--, ##STR62## α-naphthyl, β-naphthyl, furyl, benzofuryl, benzothienyl,H₂ N(CH₂)_(m) --, diphenylmethyl or ##STR63## R² is alkyl,alkyl--S(O)₀₋₂ (CH₂)_(k) S(O)₀₋₂ (CH₂)_(q) --, alkyl-O(CH₂)_(q) --, R⁵(CH₂)_(k) --O(CH₂)_(q) --, R⁵ (CH₂)_(q) --, H₂ N(CH ₂)_(q) --,cycloalkyl(CH₂)_(k) --, R¹³ CONH(CH₂)_(q) --, R¹³ NHCO(CH₂)_(q) -- or R⁶OCO(CH₂)_(q) --;R³ is --OR⁷, --NR⁷ R⁸, ##STR64## R⁴ and R¹³ areindependently hydrogen, alkyl or Y¹ --C₆ H₄ --; R⁵ is Y² --C₆ H₄ 13 , Y²--C₆ H₄ S--, Y² --C₆ H₄ O--, naphthyl, furyl, thienyl, benzofuryl,benzothienyl, indolyl or ##STR65## provided that when R⁵ is Y² --C₆ H₄S-- or Y² C₆ H₄ O--, k is 2 or 3; R¹⁴ is R⁵, mono-unsaturated loweralkyl, hydroxy, alkoxy or alkylthio, provided that when R¹⁴ is hydroxyor alkoxy, k is 2 or 3 and when R¹⁴ is mono-unsaturated alkyl oralkylthio, k is 1, 2 or 3; R⁶, R⁷ and R⁸ are independently H, alkyl,hydroxyalkyl, dihydroxyalkyl, alkoxyalkyl, dialkoxyalkyl,alkoxyalkoxyalkyl, haloalkyl, (haloalkoxy)alkyl, aminoalkyl,alkylaminoalkyl, dialkylaminoalkyl, arylalkyl or alkyl substituted witha 5-6 membered saturated ring comprising 1-2 oxygen atoms as ringmembers wherein the ring carbon atoms may be substituted with 0-2 alkylsubstituents, or R⁷ and R⁸ together with the nitrogen to which they areattached complete a 5-7 membered ring, wherein one of the 4-6 ringmembers comprising R⁷ and R⁸ may be a nitrogen atom, analkyl-substituted nitrogen atom or an oxygen atom, and wherein the ringmay be substituted on the ring carbon atoms with substituents chosenfrom alkyl and hydroxy groups; R⁹ is hydrogen, alkyl, carboxyalkyl,mercaptoalkyl, alkylthioalkyl, aminoalkyl, hydroxyalkyl, phenylalkyl,hydroxyphenylalkyl, guanidinoalkyl, imidazolyalkyl, indolylalkyl, orcarbamoylalkyl; n is 0-2; m and k are independently 0-3; q is 1-4; X andX¹ are independently a bond, --O--, --S--, or --CH₂ --; Q is hydrogen orR¹⁰ CO--; R¹⁰ is alkyl, hydroxyalkyl, alkoxyalkyl, alkylaminoalkyl,dialkylaminoalkyl, Y³ --C₆ H₄ -alkyl, alkoxy, Y³ --C₆ H₄ --, naphthyl,furyl, thienyl or pyridyl; Y, Y¹, Y² and Y³ independently represent oneor more substituents selected from H, alkyl, cycloalkyl, alkoxy, OH, F,Cl, Br, CN, --CH₂ NH₂, --CO₂ H, --CO₂ alkyl, --CONH₂ and phenyl;or apharmaceutically acceptable addition salt thereof.